Fig. 2: DNMT3A-silenced monocytes activate cardiac fibroblasts in a paracrine manner. | Nature Communications

Fig. 2: DNMT3A-silenced monocytes activate cardiac fibroblasts in a paracrine manner.

From: DNMT3A clonal hematopoiesis-driver mutations induce cardiac fibrosis by paracrine activation of fibroblasts

Fig. 2

a DNMT3A mRNA expression in PMA-activated THP-1 cells after siRNA silencing of DNMT3A normalized to RPLP0 mRNA expression (n = 4 biologically independent samples). Source data are provided as a Source Data file. b DNMT3A protein expression (isoform 1 (DNMT3A1) and 2 (DNMT3A2)) in PMA-activated THP-1 cells after siRNA silencing of DNMT3A (n = 3 biologically independent samples). Representative Western blots are shown on the left. Source data are provided as a Source Data file. c Schematic representation of the indirect co-culture experiment. d Relative ACTA2 gene expression normalized to RPLP0 mRNA expression in human cardiac fibroblasts (HCF) after incubation with supernatants from PMA-activated THP-1 cells for 48 h after siRNA silencing of DNMT3A relative or negative control (n = 5 biologically independent samples). Source data are provided as a Source Data file. e Immunofluorescence analysis of αSMA protein expression in stimulated HCF (n = 3 biologically independent experiments). Representative examples are shown left. Source data are provided as a Source Data file. f Relative TGFB1 mRNA expression normalized to RPLP0 mRNA expression analyzed by qPCR in stimulated HCF (n = 8 biologically independent samples). Source data are provided as a Source Data file. g Collagen gel contraction analysis in stimulated HCF (n = 5 biologically independent experiments) after treatment with supernatants. Source data are provided as a Source Data file. h Migration assay of stimulated HCF (n = 11 biologically independent experiments) after treatment with supernatants. Source data are provided as a Source Data file. Data are shown as mean ± SEM (a, b, d–h). Normal distribution was assessed using the Shapiro–Wilk test (a, b, d–h). Statistical analysis was performed using unpaired, two-sided Student’s t tests (a,b, d–h). t  =  5.471, 6 degrees of freedom (a); t  =  3.464, 4 degrees of freedom (b); t  =  2.490, 8 degrees of freedom (d); t  =  2.977, 4 degrees of freedom (e); t  =  2.607, 14 degrees of freedom (f); t  =  2.408, 8 degrees of freedom (g); t  =  2.562, 20 degrees of freedom (h).

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