Figure 4

Upregulation of Yy1 promotes cardiac cell cycle in dilated cardiomyopathy induced by Ttn shRNA. (A) Quantitative real-time PCR analysis of Ccnd1 and Ccnd2 expression in mouse heart tissue from control, Ttn shRNA and Yy1 treated groups. Virus does, 0.2E + 13 vg/kg. Mice were harvest four weeks after transduction. n ≥ 10. (B) Paraffin sections (left) stained with DAPI (blue), cTnI (green) and Ccnd1 (red), representing Non-CM and CM with positive Ccnd1 signal (arrowed); quantification of Ccnd1 + Non-CM and CM (right) from control, Ttn shRNA and Yy1 treated groups, n ≥ 4. (C) Paraffin section (left) and quantification (right) of Ccnd2 + in Non-CM and CM respectively, n ≥ 4. (D) Western blot and quantitative analysis (E) of Yy1, Ccnd1 and Ccnd2 protein levels in mouse heart tissue, n = 4. Data were normalized by Lamin B1 antibody. Full-length blots were presented in Supplementary Fig. S6. (F) Quantitative real-time PCR analysis of Ccnd1 and Ccnd2 expression in isolated CM from control, Ttn shRNA and Yy1 treated groups four weeks after virus transduction, n ≥ 3. (G) Paraffin section (left) and quantification (right) of EdU + in Non-CM and CM respectively, n = 4. (H) Paraffin section (left) and quantification (right) of pH3 + in non-CM, n ≥ 4. In (B), (C), (G) and (H), magnification = 100×, scale bar = 10 µm. Data were shown as mean ± SD and were normalized to total nucleus number labelled by DAPI.