Fig. 5: Myosin light chain kinase (MYLK) is a direct target of miR-1204 in vascular smooth muscle cells (VSMCs).

A, B Volcano plot showing the mRNA expression of potential miR-1204 targets in VSMCs transfected with miR-1204 mimic or miR-control (miR-ctl) combined with angiotensin II (AngII) treatment (B) or not (A). n = 3 biological replicates. Statistical analysis was performed using a two-tailed Student’s t test. C Representative plots (upper) and quantification (lower) of MLYK immunoblot analysis in VSMCs transfected with miR-1204 mimic or miR-ctl combined with AngII treatment. n = 6 biological replicates. Statistical analysis was performed using a one-way ANOVA followed by Tukey’s multiple comparison test. D Representative images (upper panel) and quantification (lower panel) of MYLK immunofluorescence staining of mouse aortas from in the indicated groups. n = 17 biological replicates for the miR-1204 + AngII group. n = 19 biological replicates for other groups. Statistical analysis was performed using a one-way ANOVA followed by Tukey’s multiple comparison test. Scale bar represents 100 μm. E Potential hsa-miR-1204 binding site in MYLK (upper panel). Luciferase activity assay confirmed that MYLK is a direct target of miR-1204 (lower panel). n = 6 biological replicates. Statistical analyses were performed using two-tailed Student’s t tests. F Representative images (left) and quantification (right) of MYLK immunofluorescent staining in aortas of healthy participants and patients with AAD. n = 6 (young normal), n = 6 (elder normal), n = 7 (young patient), n = 8 (elder patient). Statistical analysis was performed using a one-way ANOVA followed by Tukey’s multiple comparison test. Scale bar represents 20 μm. Data are presented as mean ± SD. Source data are provided as a source data file.