Fig. 3: TGFβ downregulates MYST1 in fibroblasts.
From: TGFβ promotes fibrosis by MYST1-dependent epigenetic regulation of autophagy
a mRNA (n = 4 biological replicates per group) and protein (n = 2 biological replicates per group over two independent experiments) levels of MYST1 in cultured fibroblasts stimulated with TGFβ. b Effects of siRNA-mediated knockdown of SMAD3 on mRNA (n = 5 biological replicates per group) and protein (n = 4 biological replicates per group) levels of MYST1 in TGFβ-stimulated fibroblasts. c Chromatin immunoprecipitation assay for binding of SMAD3 at three putative SMAD binding Elements (SBE) in the MYST1 promoter (n = 3 biological replicates). d Promoter assays using MYST1 promoter constructs with selective mutations at three putative SBEs, (n = 2 biological replicates for SBE1, SBE2, and SBE3 groups and n = 4 for the other groups). e Expression of MYST1 in SSc patients and healthy individuals analyzed by qRT-PCR (n = 6 patients per group) and immunofluorescence staining with Voronoi mesh-based tessellated images (n = 10 patients per group). f Expression of MYST1 in fibrotic skin of mice challenged with bleomycin or non-fibrotic control mice as analyzed by qRT-PCR (n = 4 biological replicates for control group and n = 6 for bleomycin group) and immunofluorescence staining (n = 6 biological replicates for control group and n = 8 for bleomycin group). All horizontal scale bars, 50 µm. All data are presented as median ± IQR. p-values were determined by ANOVA one-way with Sidak’s multiple comparisons test (a, c) or with Tukey’s multiple comparison post hoc test (b, d) and by two-sided Mann–Whitney test (e, f). p-values are indicated in the figure. See source data for more detailed information. rel.: relative, P4Hβ: prolyl 4-hydroxylase, SSc: systemic sclerosis, Bleo: bleomycin, fluo.: fluorescence, int,: intensity, prom.: promoter, IP: immunoprecipitation.
