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. 2020 Aug 17;130(9):4888–4905. doi: 10.1172/JCI135439

Figure 2. H19X is induced by TGF-β in a time- and dose-dependent manner in fibroblasts and related cell types.

Figure 2

Correlation of H19X expression with TGF-β pathway genes (KEGG Pathway Database). Significant correlations are highlighted in pink (Pearson P < 0.05). Genes with significant correlations and Pearson’s correlation coefficients r >│0.5│ are marked in red (A). H19X expression after 6 hours of 10 ng/mL TGF-β stimulation in SSc and HC dermal fibroblasts either in complete medium (10% FBS) or starvation medium (1% FBS) (B). Dose response curve of TGF-β–induced expression of H19X. SSc fibroblasts were stimulated with different concentrations of TGF-β (0.01–10 ng/mL) for 6 hours in 10% FBS. Time-dependent induction of H19X by TGF-β. SSc dermal fibroblasts were stimulated for 1–48 hours with 10 ng/mL TGF-β in 10% FBS medium (C). SSc fibroblasts were stimulated with PDGF-BB (20 ng/mL), IL-1β (10 ng/mL), IL-4 (10 ng/mL), IL-13 (10 ng/mL), or IL-17a (10 ng/mL) for 6 hours in 1% FBS medium (D). H19X expression in SSc fibroblasts after 6 hours of 10 ng/mL TGF-β stimulation and concomitant chemical inhibition of TGF-βR1 with SD208 (100 nM) or SB431542 (10 μM) in 10% FBS medium (E). Silencing of TGFBR2 and SMADs in SSc fibroblasts using 50 nM siRNA for a total of 54 hours and stimulated with TGF-β for the last 6 hours in 1% FBS medium (F). Correlation of H19X expression with different cell type–specific gene signatures. Significant correlations are highlighted in pink (Pearson P < 0.05). Fibroblast gene signature is marked in red (G). Stimulation of other types of fibroblasts or fibroblast-like cells with 10 ng/mL of TGF-β for 6 hours in 1% FBS medium (H). Stimulation of other cell types with 10 ng/mL of TGF-β for 6 hours in 10% FBS medium (I). H19X expression was measured by qPCR and normalized to GAPDH and RPLP0. Fold change was calculated respective to untreated control set as 1 (dashed line). Data are presented as single values and mean (n = 3–7) biological replicates. t test (B, C, H, and I) and 1-way ANOVA (D–F). *P < 0.05, **P < 0.01, ***P < 0.001.