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. 2024 Sep 28;15:8425. doi: 10.1038/s41467-024-52739-y

Fig. 4. ASPP1 facilitates cardiac fibroblast activation and collagen production in vitro.

Fig. 4

a Efficiency of small interfering RNA (siRNA) of ASPP1 in PMCFs by Western blot, n = 4 independent samples. b Effects of siASPP1 on TGF-β1-induced decreases of Col1, and Fn1 mRNA levels in PMCFs by qRT-PCR analysis, n = 6 independent samples. c, d Effects of ASPP1 knockdown (siASPP1) on TGF-β1-induced decreases of Col1 and Fn1 protein levels in PMCFs by Western blot analysis, n = 5 independent experiments. e Representative photographs of EdU-positive cells by immunofluorescent staining, n = 17 independent fields of view from three independent experiments. Scale bar = 50 μm. Magnification 20×. f Representative images of α-SMA positive cells by immunofluorescent staining and rate of α-SMA positive cells. n = 8 independent fields of view from three independent experiments in the TGF-β1 group, n = 11 independent fields of view from three independent experiments in the TGF-β1 + siASPP1 group. Scale bar = 40 μm. Magnification 40×. g Efficiency of ASPP1 overexpression plasmid in PMCFs by western blot, n = 6 independent samples. h The mRNA levels of Col1 and Fn1 in PMCFs after transfection of ASPP1 overexpression plasmids by qRT-PCR analysis, n = 6 independent samples. i, j The protein levels of Col1 and Fn1 upregulated in the PMCFs after transfection of ASPP1 overexpression plasmids by Western blot analysis, n = 4 independent experiments in Col1 group, n = 5 independent experiments in Fn1. k Representative photographs of EdU-positive cells in the ASPP1 overexpression group. n = 17 independent fields of view from three independent experiments in the NC group, n = 10 independent fields of view from three independent experiments in the oe-ASPP1 group. Scale bar = 50 μm. Magnification 20×. l Immunofluorescent staining of α-SMA and statistical analysis of α-SMA positive cells. n = 8 independent fields of view from three independent experiments. Scale bar = 40 μm. Magnification 40×. Data are represented as mean ± SEM. Statistics: Two-tailed Student’s t-test was used to calculate P values (a–d and f–l). Source data are provided as a Source data file.