Fig. 5: IRX2 is essential for fibroblast to myofibroblast transformation.
A Adult cardiac fibroblasts (CFs) were isolated from Irx2fl/fl mice and infected with an adenoviral vector carrying Cre to deplete Irx2. After 24 h of transfection, IRX2 protein expression was detected in CFs (n = 5). B Relative mRNA level of α-Sma in Irx2-depleted adult mouse CFs (n = 5). C Representative images and quantification of α-SMA (green) in Cre-infected Irx2fl/fl CFs in response to Ang II incubation for 24 h. Nuclei are stained with DAPI (blue). (n = 5, for each group, 50-60 fields were counted). D CFs were isolated from wild-type mice and then transfected with shRNAs targeting Irx2. After 24 h of transfection, the cells were subjected to Ang II treatment for 24 h to induce a fibrotic phenotype. After that, the cells were harvested for the detection of α-SMA with western blotting (n = 4). E Relative mRNA level of Col1 in IRX2-deficient CFs after Ang II incubation for 24 h (n = 4). F CFs were isolated from wild-type mice and then overexpressed Irx2 with an adenovirus vector, after which they were subjected to Ang II treatment for 24 h. The relative mRNA level of α-Sma was detected to reflect fibroblast to myofibroblast transformation (n = 5). G CFs were isolated from wild-type mice and then overexpressed Irx2 with an adenovirus vector, after which they were subjected to Ang II treatment for 24 h. Total RNA was extracted, and RNA sequencing was performed. Volcano plots indicating the differentially expressed genes (DEGs) in Irx2-overexpressing CFs relative to controls after Ang II administration for 24 h. H Heatmap of several differentially expressed genes in Irx2-overexpressing CFs relative to controls after Ang II infusion. Data are shown as the mean ± SEM, and analysed using one-way ANOVA followed by Tukey post hoc test (B and D–F) or Tamhane’s T2 test (C). For the analysis in (A), an unpaired two-tailed Student′s t test was conducted. Source data are provided as a Source Data file.
