Fig. 5: miR-203a-3p targets PCSK1. | Nature Communications

Fig. 5: miR-203a-3p targets PCSK1.

From: Epigenetic regulation of beta-endorphin synthesis in hypothalamic arcuate nucleus neurons modulates neuropathic pain in a rodent pain model

Fig. 5

a Venn diagrams showing thirteen putative target genes of miR-203a-3p in the intersection predicted by the combined use of the “TargetScan” (yellow), miRDB” (blue), “miRanda” (green) and “miRwalk” (pink) algorithms. b qPCR analysis indicating that six of the predicted target genes are downregulated by more than 50% in the ARC of rats at Day 14 post-CCI-ION-operation. **p < 0.01, ***p < 0.001 versus sham, two-sided Student’s t-test. n = 8 rats for each group. c Diagrams illustrate luciferase reporters containing wild-type (PCSK1-wt) or mutant PCSK1 (PCSK1-mut). d The luciferase reporter assay shows that miR-203a-3p interacts with the 3’-UTR of PCSK1. ***p = 0.0005 versus miR-203a NC, one-way ANOVA followed by Bonferroni’s test. Data are from six independent experiments. e Diagrams illustrate sequences of PCSK1 3’-UTRs with highly conserved miR-203a-3p-binding sites in various vertebrates. f FISH analysis of the DIG-labeled miR-203a-3p probe (red) combined with immunofluorescent labeling with PC1 (green) in ARC sections 14 days after CCI-ION or sham surgery. Arrows show colocalization. n = 4 rats per group. Scale bar = 25 μm. g Bar charts show the percentage of double-stained ARC neurons among total numbers of miR-203a-3p+- or PC1+-labeled neurons. ***p = 0.0002 versus miR-203a + sham, ##p = 0.0019 versus miR-203a + CCI-ION, two-sided Student’s t test. n = 4 rats per group. h, i Representative immunoblots and bar charts showing that intra-ARC injection of agomir-203a (h) or miR-203a-up (i) decreased PC1 expression in the ARC of intact rats. **p = 0.0039 versus miR-NC, ***p = 0.0002 versus agomir-NC, one-way ANOVA followed by Bonferroni’s test. n = 4 rats per group. All data are presented as mean values ± SEM.

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