Fig. 3.

Knockdown or inhibition of Dot1L attenuates Ang II-induced fibrosis in neonatal rat CFs (NRCFs). A Dot1L, CTGF, FN, and H3K79me3/2/1 protein expression level in NRCFs stimulated by Ang II (2 μmol/L) for different time (0, 6, 12, 24, 36, and 48 h). B, C Knockdown of Dot1L attenuates ECM deposition in Ang II-treated NRCFs. Dot1L siRNA and control siRNA were transfected into NRCFs, followed by Ang II stimulation for 48 h. The protein expressions of Dot1L, Col 1, Col 3, CTGF, and H3K79me3 were analyzed by western blot (B); Representative images of immunofluorescence staining of Dot1L and Col 1 (C). D EPZ5676 attenuates ECM deposition in Ang II-treated NRCFs. NRCFs were pretreated with 10 and 20 μmol/L EPZ5676 for 4 h and then co-incubated with 2 μmol/L Ang II for 48 h. CTGF, Col 3, Col 1, FN, and H3K79me3 were analyzed by western blot. All data are presented as mean ± SD. ANOVA, ^*p < 0.05, ^***p < 0.001 versus Ctrl or Ctrl + siCtrl, ^#p < 0.05, ^###p < 0.001 versus Ang II or Ang II + siCtrl, each acquired from three individual experiments