Fig. 3: MARK4 and AGTR1 are co-targets of miR199a-5p.

a The volcano plot displaying the differential expressed genes in cardiomyocytes transfected with miR199a-5p or NC mimic. Red and green colors indicate up-regulated or down-regulated genes. b qPCR of top 10 candidate target genes in miR199a-5p, NC mimic or antimiRNA treated NRVMs in vitro, n = 3 independent sample. c Luciferase vectors containing the miR199a-5p-binding sites of AGTR1 were delivered to 293 T cells with or without co-delivery of miR199a-5p. Twenty-four hours after the transfection, luciferase activity was measured, n  =  4 independent samples. d The cardiomyocytes were transfected with either miR199a-5p or antimiR199a-5p for 72 h. The expression levels of AGTR1 were examined by western blotting, n  =  4 independent samples. e Bubble chart showing the enrichment result of biological process between miR199a-5p or negative control (NC) mimic transfected cardiomyocytes analyzed by KEGG pathway. f Predictive analysis of miR199a-5p key target genes by miRBD, miRmap, tragetscan, and microT site. The number in the Venn diagram indicates the number of miRNAs belonging to the corresponding intersection. g Luciferase assays confirmed that miR199a-5p is associated with the MARK4 mRNA 3 ’ -UTR. Constructs (vectors) carrying or not carrying the MARK4 3′- UTR were cotransfected with scramble NC mimics or miR199a-5p mimics in 293 T cells, n = 3 independent sample. h Western blot analysis of MARK4 protein expression in cardiomyocytes treated with NC, miR199a-5p or antimiRNA, n  =  4 independent samples. Quantitative data were expressed as the mean ± SD. of at least 3 independent experiments. The p-values for Fig. 2d and h were generated using an unpaired two-tailed Student’s t-test, p value for Fig. 2b, c, g were generated by one‐way analysis of variance (ANOVA), followed by Tukey’s multiple‐comparison post hoc test. *P < 0.05, **P < 0.01, NS, not significant. Source data are provided as a Source Data file.