Fig. 5
From: Histone lysine dimethyl-demethylase KDM3A controls pathological cardiac hypertrophy and fibrosis
KDM3A activates the transcription of Timp1 in cardiomyocytes. Cardiomyocytes (CM) and cardiac fibroblasts (cFb) were isolated from WT and Kdm3a-Tg mouse hearts at week 6 post-Sham and TAC surgery. a–e Transcripts of fetal genes and fibrosis-related ECM genes were measured by qRT-PCR, normalized against internal Gapdh, and expressed relative to WT Sham CM. n = 3 ± SEM, *, #p < 0.05 by student t test. *, WT TAC vs WT Sham. #, Tg-TAC vs WT-TAC. f Cardiomyocytes from Kdm3a-Tg mouse hearts at week 6 post-Sham and TAC surgery were isolated and used for ChIP assay with antibodies against KDM3A (left panel) or H3K9me2 (right panel). The relative occupancies of KDM3A and H3K9me2 at Timp1 promoter were normalized against Input and expressed relative to Sham control. n = 3 ± SEM, *p < 0.05 (t test). g Timp1-Luciferase reporter was transfected in combination with plasmids as indicate into 293 T cells. Cells were harvested 48 h after transfection. Luciferase activities were normalized against co-transfected β-galactosidase, expressed relative to vector-transfected cells. n = 3 ± SEM. (h-k) Kdm3a-Tg mice were transduced with adenovirus AAV9 expressing control or Timp1 shRNA and subjected to Sham or TAC surgery. Hearts were echoed and harvested 5 weeks post-surgery. h WB of TIMP1 and GAPDH, i HW/BW, j %FS, and k percent of fibrotic area of Sham (S), TAC (T) Kdm3a-Tg mice transduced with control or Timp1 shRNA. n = 7 ± SEM, *p < 0.05 (ANOVA)
