Figure 2: H3K4me2 editing induces loss of SMC contractility in vitro.

A. Transcript expression of contractile genes in Myocd-LSD1 and Myocd-LSD1^NF SMC. Expressed as fold change compared to control SMC. B. ACTA2 and DAPI immunofluorescent staining in Myocd-LSD1 and Myocd-LSD1^NF SMC. Scale bar = 100 μm. C. SMC-related gene protein expression in control, Myocd-LSD1/LSD1^NF SMC. D. Protein expression quantification normalized to GAPDH. E. Collagen contraction assay using control, Myocd-LSD1 and Myocd-LSD1^NF SMC. Scale bar = 1mm. Measurement of diameter over time. F. Ex-vivo wire myography combined with potassium chloride infusion in thoracodorsal artery rings incubated with Myocd-LSD1 or control lentivirus for 48h (n=4 mice per group). G. mRNA expression of myocardin, Acta2 and Tagln in Myocd-LSD1 and Myocd-LSD1^NF SMC transduced with Adv-GFP or Adv-Myocardin. H. Acta2 transcript expression in Myocd-LSD1 and Myocd-LSD1^NF SMC treated with rapamycin (100 nM). I. Collagen contraction assay using control, Myocd-LSD1^NF, Myocd-LSD1 and Myocd-LSD1 SMC treated with rapamycin (100 nM). Gel area variation at day 5 post-gelation normalized to baseline area. Scale bar = 1 mm. Data are represented as mean ± s.e.m of 3-6 independent experiments. Groups were compared by Student t-test, One-way ANOVA, Two-way ANOVA, or Mann-Whitney (F). * p<0.05, ** p<0.001, *** p<0.0001.