E2F transcription factor 1 (E2F1) promotes the transforming growth factor TGF-β1 induced human cardiac fibroblasts differentiation through promoting the transcription of CCNE2 gene

Figure 1.. E2F1 was highly expressed in HCFs induced by TGF-β1. (a, b) Differentiation associated proteins (a) and mRNAs (b) in TGF-β1 stimulated HCFs in manner of time (loading control: GAPDH). n = 3 *p < 0.05, **p < 0.01, ***p < 0.005 ****p < 0.001 vs 0 h. C The plates that cells were cultured exerted no significant impact on differentiation. D, E Timeline of E2F1, E2F2, E2F3 protein (d) and mRNA (e) expression after TGF-β1 stimulating in manner of time. n = 3. *p < 0.05, **p < 0.01. ***p < 0.005. ****p < 0.001 vs 0 h

Figure 2.. Silenced the expression of E2F1 demoted the differentiation of HCFs. (a, b) Differentiation associated proteins (a) and mRNAs (b) expression in TGF-β1 induced HCFs (loading control: GAPDH). n = 3. C-E migration (c, d) and proliferation (e) abilities of HCFs. n = 5. *p < 0.05, **p < 0.01, ***p < 0.005 ****p < 0.001

Figure 3.. E2F1 promoted the transcription of CCNE2. (a) KEGG showed that CCNE family was key mediators in cell proliferation. (b, c) Construction of CCNE2 WT and MUTANT promoters (c) and pmirGLO vectors (b). (d) Luciferase activity in E2F1 pcDNA3.1 + CCNE2 mutant promoter. n = 4. (e, f) Expression of protein (e) and mRNA (f) of CCNE2 in TGF-β1 stimulated HCFs (loading control: GAPDH). n = 3. G Immunoprecipitation of anti-E2F1 to CCNE2 in HCFs lysates. *p < 0.05, **p < 0.01

Figure 4.. E2F1 promoted the TGF-β1 induced HCFs differentiation through promoting the transcription of CCNE2. (a) Differentiation associated proteins expression in TGF-β1 induced HCFs (loading control: GAPDH). n = 3. B-D Migratory (b, c) and proliferative (d) abilities of HCFs. n = 5. *p < 0.05, **p < 0.01, ***p < 0.005 ****p < 0.001