Fig. 6: MEG3 inhibits VSMC phenotypic switching via increasing R-Ras2.

a Volcano plot of deregulated transcripts in HASMCs stimulated with Ang II compared with PBS. The p-value was calculated by a Wilcoxon rank sum test. No correction was applied for multiple testing. b MEG3 expression level in aorta of AAA patients and healthy control individuals (non-AAA). n = 6 distinct samples for each group, two-sided Unpaired t-text. c Meg3 expression level in aorta of Sham and AAA mice. n = 5 mice for each group, two-sided Mann–Whitney test. d Fluorescence in situ hybridization (FISH) assay to determine the expression of MEG3 in aortic sections from AAA patients and non-AAA. Scale bar: 50 μm. MEG3: Red, DAPI: Blue. 3 independent experiments showed consistent results. e Fluorescence in situ hybridization (FISH) assay to determine the expression of MEG3 in HASMCs simulated with Ang II. Scale bar: 50 μm. MEG3: Red, DAPI: Blue. 3 independent experiments showed consistent results. f Representative western blotting and quantification of R-Ras2 in HASMCs transfected with si-MEG3. n = 6 distinct samples for each group, two-sided unpaired t-test. g Representative western blotting and quantification of R-Ras2 in HASMCs overexpressing MEG3 and stimulated with Ang II. n = 6 distinct samples for each group. Two-way ANOVA test followed by Turkey’s multiple comparisons test. h HASMCs were overexpressed with MEG3, followed by transfected with si-RRAS2 and stimulated with Ang II. Protein levels of Calponin 1, α-SMA, SM22, and R-Ras2 were determined and their relative expression levels were analyzed. n = 6 distinct samples for each group. Two-way ANOVA test followed by Turkey’s multiple comparisons test. i Representative image of in situ zymography for detection of MMPs activity (Green). DAPI: Blue. Scale bar = 50 μm. n = 6 distinct samples for each group. Two-way ANOVA test followed by Turkey’s multiple comparisons test. All data were represented as mean ± SEM. Source data are provided as a Source Data file.