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11 publications mentioning ssc-mir-34c-2

Open access articles that are associated with the species Sus scrofa and mention the gene name mir-34c-2. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary.

1
[+] score: 310
The full-length blot images are presented in Supplementary Figure  3. Reciprocal regulation between Notch1 and miR-34cAs overexpressing N1ICD decreases miR-34c expression during PSCs development (Fig.   1) and miR-34c mimics decreases Notch1 gene expression in the proliferation period (Fig.   2), differentiation day 1and differentiation day 7 (Fig.   4), these results suggest a regulatory feedback may exist between Notch1 and miR-34c. [score:10]
The full-length blot images are presented in Supplementary Figure  3. As overexpressing N1ICD decreases miR-34c expression during PSCs development (Fig.   1) and miR-34c mimics decreases Notch1 gene expression in the proliferation period (Fig.   2), differentiation day 1and differentiation day 7 (Fig.   4), these results suggest a regulatory feedback may exist between Notch1 and miR-34c. [score:9]
Since our in vitro study has shown that overexpressing miR-34 inhibits muscle development, we believe the miR-34c overexpression experiment alone would be sufficient to demonstrate the role of miR-34c in vivo. [score:8]
Overexpressing N1ICD decreases miR-34c expression in vitroTo explore the regulations of Notch1 signaling in PSCs development, we constructed the constitutively activated N1ICD PSCs. [score:7]
Our study ascertains that miR-34c inhibits PSCs proliferation by inhibiting Notch1 expression. [score:7]
These results demonstrate we have successfully overexpressed N1ICD in PSCs and N1ICD decreased miR-34c expression in all three periods of PSCs development. [score:6]
Figure 1Overexpressing N1ICD decreases miR-34c expression during PSCs development. [score:6]
But miR-34c inhibitor had no effect on Myod gene expression on differentiation day 7 (Fig.   5D and E). [score:5]
Figure 2Overexpressing miR-34c mimics inhibits PSCs proliferation. [score:5]
The full-length blot images are presented in Supplementary Figure  3. Next, miR-34c inhibitor or Control (miR-34c inhibitor and Control are both synthetic oligonucleotide sequences; see Table  S1 for details) was transfected into PSCs, and the cells were induced to differentiation for 1 day and 7 days. [score:5]
Overexpressing miR-34c inhibits PSCs proliferation in vitro. [score:5]
Our results not only established Notch1 is the target gene of miR-34c but also discovered that Notch1, in turn, directly regulates miR-34c in PSCs. [score:5]
Figure 3Overexpressing miR-34c inhibitor increase PSCs proliferation. [score:5]
But miR-34c inhibited the skeletal muscle satellite cell proliferation by targeting Notch1, and miR-34c reduced the number of satellite cells to be fused to existing myofibers, resulting in smaller muscle fiber diameter after miR-34c injection. [score:5]
In this study, we used miR-34c mimics and inhibitor to manipulate the miR-34c level in transfected PSCs and that elevated miR-34c not only inhibits PSCs proliferation but also promotes PSCs differentiation. [score:5]
The full-length blot images are presented in Supplementary Figure  3. Next, miR-34c inhibitor or Control (miR-34c inhibitor and Control are both synthetic oligonucleotide sequences; see Table  S1 for details) was transfected into PSCs, and the cells were induced to differentiation for 1 day and 7 days. [score:5]
Figure 5Overexpressing miR-34c inhibitor reduces PSCs differentiation. [score:5]
The expression of miR-34c was decreased in the N1ICD overexpressed cells (p < 0.01; Fig.   1C). [score:5]
From the RNA-seq data, we discovered that many miRNAs were differently expressed in the N1ICD overexpressing PSCs, including miR-34c [29]. [score:5]
Since N1ICD expression was decreased after PSCs transfected with miR-34c mimics (Figs  2 C and 4B), Notch1 may be a potential target gene of miR-34c. [score:5]
Overexpressing N1ICD decreases miR-34c expression in vitro. [score:5]
For all the experimental data in this study, including miR-34c inhibiting PSCs proliferation, miR-34c promoting PSCs differentiation and miR-34c repressing mice muscle development, we are comparing only the control group to the treatment group; thus we used the student’s t-test for the statistical analyses (SPSS 18.0, Chicago, IL, USA). [score:4]
In conclusion, our research demonstrates that miR-34c inhibits PSCs proliferation but promotes PSCs differentiation in vitro, and miR-34c represses pig muscle development in vivo. [score:4]
These results suggest that Notch1 is the direct target gene of miR-34c. [score:4]
However, there is no report regarding the role of miR-34c on PSCs development; therefore, the objective of this study was to define the role of miR-34c on PSCs development and ascertain whether there is a regulatory relationship between miR-34c and N1ICD. [score:4]
This reciprocal regulatory loop formed by miR-34c and Notch1 that controls skeletal muscle development is novel, and this information expands our understanding of the mechanisms involved in muscle development. [score:4]
To establish N1ICD regulates miR-34c expression, N1ICD was transfected into PSCs in primary culture. [score:4]
Furthermore, the objective of this particular experiment was to ascertain whether the miR-34c inhibits muscle development in vivo. [score:4]
Taken together, these results demonstrate miR-34c inhibits PSCs proliferation in vitro. [score:3]
Another miR-34 family member miR-34c also has been shown to inhibit rat vascular smooth muscle cell proliferation [27]. [score:3]
This study would be better to have the miR-34c inhibitor group in the in vivo study. [score:3]
PSCs transfected with pCDNA3.1- N1ICD, miR-34c inhibitor, miR-34c mimics or Control by Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA), according to the manufacturer’s instructions. [score:3]
We have also established Notch1 is a direct target gene of miR-34c using a dual-luciferase reporter assay. [score:3]
These results demonstrate there exists a regulatory loop between Notch1 and miR-34c in PSCs development. [score:3]
However, after considerable deliberations, we decided to forgo the miR-34c inhibitor group for the following reason: the mice used in this study are 4-week-old. [score:3]
Representative images of immunofluorescent staining of PSCs transfected with miR-34c inhibitor. [score:3]
The mRNA level of myogenin, myosin, and MyHC was elevated by the miR-34c overexpression on differentiation day 7 (p < 0.01, Fig.   4C), and protein level of myogenin and myosin follows the same pattern as the mRNA (p < 0.05, Fig.   4D). [score:3]
PSCs transfected with miR-34c mimics, miR-34c inhibitor or Control and maintained in growth medium. [score:3]
However, on differentiation day 7, overexpression of miR-34c showed no significant influence on MyoD mRNA and protein levels (Fig.   4C and D). [score:3]
Figure 4Overexpressing miR-34c mimics promotes PSCs differentiation. [score:3]
The full-length blot images are presented in Supplementary Figure  2. Next, we used miR-34c inhibitor to further ascertain the role of miR-34c on PSCs proliferation. [score:3]
Therefore, injection of LV-miR-34c inhibitor may not obtain additional muscle growth; thus, the value of using more mice may be questionable following the principle of a minimal number of animals to be used in any experiments. [score:3]
Overexpressing miR-34c reduced the percentage of Edu positive cells (p < 0.05, Fig.   2A). [score:3]
The full-length blot images are presented in Supplementary Figure  2. Next, we used miR-34c inhibitor to further ascertain the role of miR-34c on PSCs proliferation. [score:3]
MiR-34c inhibitor reduced the endogenous miR-34c level (Fig.   3B). [score:3]
But miR-34c inhibitor had no effect (p > 0.05) on myogenin and MyHC mRNA level on differentiation day 1 (Fig.   5A). [score:3]
Overexpression of miR-34c reduced Notch1 mRNA, and elevated MyoD mRNA and myosin mRNA on differentiation day 1 (p < 0.05, Fig.   4A). [score:3]
PSCs transfected with miR-34c inhibitor increased the mRNA and protein levels of N1ICD on both differentiation day 1 and day 7 (Fig.   5). [score:3]
On differentiation day 7, western blot result shown miR-34c inhibitor decreased (p < 0.05) myogenin and myosin protein levels. [score:3]
The full-length blot images are presented in Supplementary Figure  1. Overexpressing miR-34c inhibits PSCs proliferation in vitroFirst, we investigated the role of miR-34 in PSCs proliferation. [score:3]
After transfected with miR-34c inhibitor, the mRNA level of Notch1, CCNB, CCND and CCNE and PCNA were increased and p21 was decreased (p < 0.05, Fig.   3B). [score:3]
As for protein level of these genes, western blot result showed overexpression of miR-34c reduced Notch1, and elevated MyoD and myogenin on differentiation day 1 (p < 0.05, Fig.   4B). [score:3]
On differentiation day 1, western blot result shown miR-34c inhibitor decreased (p < 0.05) MyoD and myosin protein levels, but had no effect on myogenin (Fig.   5B). [score:3]
Overexpressing miR-34c promotes PSCs differentiation. [score:3]
We injected miR-34c into the gastrocnemius muscle of the mice to establish the changes in muscle growth and gene expressions. [score:3]
We measured the expression of Notch1 and the myogenic marker genes after PSCs were transfected with miR-34c inhibitor. [score:3]
Finally, through miR-34c lentivirus injection into mice gastrocnemius muscle, we confirmed miR-34c represses mice muscle development. [score:2]
Using, we showed that CSL-N1ICD complex binds directly to the −3631~−3625 sites of miR-34c. [score:2]
As the miR-34c level is reduced when N1ICD was overexpressed during PSCs development (Fig.   1A) and a CSL-N1ICD complex binding site (GTGGGAA) exists at upstream of the miR-34c genomic site (Fig.   6C), we measured the DNA fragment binding with CSL-N1ICD complex by ChIP. [score:2]
MiR-34c mimics and inhibitor were (see Table  S1) purchased from GENEWIZ (Suzhou, China). [score:2]
But, the role of miR-34 plays in pig skeletal muscle development has not been reported. [score:2]
These findings establish there exists a regulatory loop between Notch1 and miR-34c. [score:2]
These results demonstrate that injecting LV-miR-34c miR-34c represses muscle development in vivo. [score:2]
MiR-34c inhibitor decreased MyoD and myosin mRNA level on differentiation day 1 (p < 0.05, Fig.   5A). [score:2]
Through the dual-luciferase reporter assay, we found N1ICD decreased the pGL3-basic-miR-34 upstream recombinant vector relative luciferase activity, but this inhibition was abolished by the mutated CSL-N1ICD complex binding site (GTGGGAA) (Fig.   6F). [score:2]
Reciprocal regulation between Notch1 and miR-34c. [score:2]
Consistent with this result is the average area of myofibers decreased (p < 0.01, Fig.   7C), which means miR-34c repressed muscle development in vivo. [score:2]
This result indicates CSL-N1ICD complex may regulate miR-34c transcription. [score:2]
In our study, we used Edu assay to confirm miR-34c inhibits PSCs proliferation, and our qRT-PCR and western blot results show miR-34c is positively correlated with p21, and negatively correlated with CCNB, CCND, and CCNE. [score:2]
Thus the presence of virus affected muscle development, which explains the gastrocnemius muscle weight of the injected virus groups (LV-Control and LV-miR-34c) was lower than that of the Normal group. [score:2]
MiR-34c has been shown to be a new modulator of VSMC proliferation through targeting SCF [27]. [score:2]
MiR-34c inhibitor increased the percentage of Edu positive cells (p < 0.01, Fig.   3A). [score:2]
The CSL-N1ICD complex binds directly to the -3631~-3625 sites of miR-34c. [score:2]
MiR-34c represses muscle development in vivoTo evaluate the function of miR-34c in vivo, we injected lentivirus expressing miR-34c mimics or Control into mice gastrocnemius muscle. [score:2]
To our knowledge, there is no report about the miR-34c function on skeletal muscle development. [score:2]
MiR-34c inhibitor decreased (p < 0.05) myogenin, myosin and MyHC genes mRNA level on differentiation day 7 (Fig.   5D). [score:2]
In human, three miR-34 precursors are produced from two transcriptional units, miR-34a precursor is transcribed from chromosome 1, and miR-34b and miR-34c precursors are co-transcribed from a region on chromosome 11 [23]. [score:1]
The miR-34 family members (miR-34a, miR-34b, and miR-34c) were discovered computationally [20] and later verified by experiment 21, 22. [score:1]
Either pGL3-basic-miR-34c upstream-mut or pGL3-control was used as a control for pGL3-basic-miR-34c upstream. [score:1]
But only miR-34a and miR-34c are found in pig 24, 25. [score:1]
Thus, to ascertain the miR-34c function observed in our cell culture studies, we conducted the in vivo study in mice. [score:1]
To evaluate the function of miR-34c in vivo, we injected lentivirus expressing miR-34c mimics or Control into mice gastrocnemius muscle. [score:1]
We found relative luciferase activity was decreased (p < 0.01; Fig.   6B) when HEK-293T cells were co -transfected with miR-34c mimics and pmirGLO- Notch1-3′UTR. [score:1]
Mice were purchased from Guangdong Medical Lab Animal Center, and lentivirus containing miR-34c mimics or Control were purchased from Shanghai JiKai Gene Chemical Technology Co. [score:1]
MiR-34c or microRNA-control was delivered by a lentiviral vector (LV). [score:1]
We measured the expression of Notch1 and the myogenic marker genes after PSCs were transfected with miR-34c mimics. [score:1]
As shown in Fig.   6E the miR-34 upstream of its genomic site (about 4600 bp) is inserted into the pGL3-basic vector. [score:1]
Body weights were no difference among all groups, but the weights of gastrocnemius muscle were decreased in the LV-miR-34c treatment group and Normal groups (p < 0.05, Fig.   7D). [score:1]
*Indicates a difference between Control and miR-34c mimics. [score:1]
However, miR-34c mimics had no effect on CCNB mRNA and protein levels (Fig.   2B and C). [score:1]
Western blot confirmed that miR-34c mimics reduced (p < 0.05) Notch1, CCND and CCNE protein level, and increased (p < 0.01) p21 protein level (Fig.   2C). [score:1]
So we constructed the pGL3-basic-miR-34 upstream recombinant vector (pGL3-basic-miR-34 upstream). [score:1]
The miR-34c mimics decreased Notch1, CCND, CCNE and PCNA mRNA levels and increased p21 mRNA level (p < 0.05, Fig.   2B). [score:1]
A ~4600 bp sequence upstream of miR-34c in genomic DNA was amplified and inserted into pGL3-basic Vector (Ambion, Carlsbad, CA, USA). [score:1]
qRT-PCR result showed miR-34c mimics increased (p < 0.001) cellular miR-34c level at 24 h after transfection (Fig.   2B). [score:1]
Figure 6Negative feedback between miR-34c and Notch1. [score:1]
Total fibers in one field were increased after injection with LV-miR-34c. [score:1]
CCND1 mRNA level was decreased after LV-miR-34c injection, but mRNA levels of CCNB1 and p21 were increased (p < 0.05, Fig.   7E). [score:1]
MiR-34c represses muscle development in vivo. [score:1]
After injecting LV-miR-34c, MyoD protein level was decreased (Fig.   7F) and myogenin protein level was increased (Fig.   7F), but their mRNA levels were not different (Fig.   7E). [score:1]
Mice were injected with physiological saline, LV-Control or LV-miR-34c. [score:1]
Both miR-34c mimics and Control are synthetic oligonucleotide sequences delivered by the lentiviral vector. [score:1]
N1ICD with pGL3-basic-miR-34 upstream recombinant vector relative or pGL3-basic-miR-34 upstream (mut) recombinant vector were transfected into HEK-293T cells respectively. [score:1]
Notch1 mRNA level was decreased in the LV-miR-34c injection group (p < 0.05, Fig.   7F). [score:1]
Using dual-luciferase reporter assay and Chromatin immunoprecipitation (ChIP), we demonstrated there is a regulatory loop between Notch1 and miR-34c. [score:1]
These results indicate elevated miR-34c promotes PSCs differentiation. [score:1]
A fragment about 400 bp long located at -3631 upstream of miR-34c was amplified (Fig.   6D). [score:1]
LV-miR-34c injection also increased myosin mRNA level (p < 0.05, Fig.   7E). [score:1]
Lanes 1–7 represent DL 10000, pGL3-basic, double digested pGL3-basic, double digested miR-34 upstream of its genomic site, pGL3-basic-miR-34 upstream recombinant vector, double digested pGL3-basic-miR-34 upstream recombinant vector, DL5000, respectively. [score:1]
The full-length blot images are presented in Supplementary Figure  2. The medium was changed to differentiation medium for 1 and 7 days to study the function of miR-34c on PSCs differentiation. [score:1]
In Fig.   6D *Indicates differences between Normal and LV-Control or Normal and LV-miR-34c in mice gastrocnemius muscle. [score:1]
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2
[+] score: 28
Previous studies showed that miR-34c alone could not induce the entire male reproductive process but still plays an important role in the differentiation of murine embryonic stem cells into male germ cells by targeting the retinoic acid receptor gamma gene (RARg) 49 and in germ cells by being highly expressed in spermatocytes and round spermatids; miR-34c over -expression lead to down-regulation of the TGIF2 gene, an inhibitor of the TGF-β pathway which is crucial for spermatogenesis 17. [score:12]
In both pig breeds, 14 miRNAs were identically up-regulated after sexual maturity (Supplementary Table S14); the relative expression fold-change of miR-145-5p, miR-205, miR-34c, miR-9, miR-9-1 and miR-9-2 at 200 days old were more notable, especially that of miR-34c, which were consistent with the findings of Lian 30 (Fig. 3). [score:6]
Before sexual maturity, only one DE miRNA (i. e. miR-34c) was down-regulated (Supplementary Table S14). [score:4]
Among these DE miRNAs, 14 were co-expressed, including miR-218-5p and miR-34c. [score:3]
In addition, in the inter-breed analysis, miR-34c showed a significantly lower expression in LW pigs than in LC pigs at 20 days. [score:3]
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3
[+] score: 19
This pattern included the two created target sites for ssc-miR-34a and ssc-miR-34c, both predicted by TargetScan and PACMIT in SLA-1 (Figure 3 A); the disrupted target site for ssc-miR-148a in HSPA1A predicted by PACMIT and TargetSpy (Figure 3 B); the ssc-miR-133b (TargetScan and PACMIT), ssc-miR-133a-3p (TargetScan) and ssc-miR-323 (TargetSpy) created target sites in RNF5 (Figure 3 C); and the disrupted site for ssc-miR-2320 predicted by TargetSpy in SLA-1 (Figure 3D). [score:19]
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4
[+] score: 14
Two of the additional differentially expressed miRNAs from the sequencing data, mir-34c and mir-489 could not be analyzed by qPCR due to very low expression in the majority of the samples. [score:5]
In other studies mir-34c has been shown to be expressed during adipogenesis of 3T3-L1 adipocytes and in murine brown adipocytes. [score:3]
Also mir-489 and mir-34c could not be detected in all animals due to very low expression, and were therefore excluded from the qPCR analysis. [score:3]
Differential expression in the sequencing data from the lean and obese pigs was calculated using the DESeq2 package in R and revealed six significantly differentially expressed miRNAs between the two groups: mir-9-5p, mir-124a-3p, mir-9-3p, mir-199a-5p, mir-489-3p and mir-34c-3p. [score:3]
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5
[+] score: 13
The over -expression of miR-34c in HeLa cells led to a shift of the expression profile toward the germinal lineage, and miR-34c could play a role in the late steps of spermatogenesis [12]. [score:5]
One of these putative target genes is deleted in azoospermia like gene (DAZL), targeted by miR-34b, miR-34c etc. [score:5]
Five miRNAs (mmu-miR-449, rno-miR-34b, mmu-miR-34c hsa-miR-181d, mmu-miR-214) appeared to be differentially expressed in two independent reports [18], [19]. [score:3]
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6
[+] score: 10
Secondly, targets for a sub-set of down-regulated miR (miR-15, miR-16, miR-27, miR-29, miR-34 and miR-106), of which 44 targets were identified based on our previous criteria were predicted (see Additional file 4). [score:8]
These miR have been implicated in multiple cellular processes including cell growth (miR-24), apoptosis (miR-16, miR-24 and miR-29), and cell cycle regulation in normal (miR-16) and cancerous cells (miR-24, miR-27, miR-29 and miR-34) [9, 40- 46]. [score:2]
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7
[+] score: 7
Other miRNAs from this paper: ssc-mir-34a, ssc-mir-34c-1
For example, miR-34c inhibits lung cancer proliferation, migration and invasion by targeting PDGFRα/β [19]; miR-34a affects the growth of pulmonary artery smooth muscle cells in human by targeting PDGFRα [20]. [score:7]
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8
[+] score: 6
In addition to tissue-specific ageing, it is increasingly evident that many miRNA regulate gene expressions in well-known ageing pathways, most notably in the p53 tumor suppressor pathway (miR-34, miR-29 and miR-217, etc. ) [score:6]
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9
[+] score: 4
Among these, miR-363, miR-423 and miR-34 were the top three upregulated miRNAs in Duroc pigs with fold change ranging from 3.29 to 1.68. [score:4]
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10
[+] score: 2
These include ssc-miR-7 [56], ssc-miR-15a [26], ssc-miR-18a [26, 56], ssc-miR-21 [55, 56], mmu-miR-34b-3p [26], ssc-miR-34c [26], ssc-miR-92b-3p [26], ssc-miR-193a-3p [55], hsa-miR-449a [26], and ssc-miR-671-3p [26]. [score:1]
Only two miRNAs were significantly changed throughout the whole experiment: ssc-miR-34c and ssc-miR-92b-3p. [score:1]
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11
[+] score: 1
Many immune-related miRNAs have been identified in innate and adaptive immune systems, including the miR-17—92 cluster, miR-221, miR-10, miR-196b, miR-126, miR-155, miR-150; miR-181a, miR-326, miR-142-3p, miR-424, miR-21, miR-106a, miR-223, miR-146; the let-7 family, miR-9, and miR-34 [6]. [score:1]
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