Fig 2: BMP4 reduced liver fibrogenic and inflammatory markers in TGF-β1-induced 3D spheroids.a, Representative immunofluorescence images of 3D spheroids (IHH/LX-2, 24:1), treated with TGF-β1 in presence or absence of BMP4 or GREM1 for 48 h, stained for COL1A1 (green), αSMA (red) and nuclei (DAPI, blue). Scale bars represent 50 µm. b, Bar graph displays fluorescence intensities of COL1A1 and αSMA quantified using ImageJ and normalized to the number of nuclei (n = 15–30 spheroids from three different experiments). c, RT–qPCR analysis of COL1A1, αSMA, IL-8 and CTGF in 3D spheroids, treated with TGF-β1 in presence or absence of BMP4 (n = 4 biologically independent experiments). d, RT–qPCR analysis of IL-8, CTGF and COL1A1 in TGF-β1-treated 3D spheroids, stimulated with BMP4 in presence or absence of GREM1 (n = 3 biologically independent experiments). Values are mean ± s.e.m. Statistical significance was determined using one-way ANOVA followed by Bonferroni post hoc test or Kruskal–Wallis with post hoc Dunn’s test.Source data

Fig 3: Hepatic senescence markers are associated with hepatic fibrosis.a, RT–qPCR analysis of hepatic fibrosis markers (TGFβ1, COL1A1 and αSMA) in lean individuals, patients with NAFLD and patients with NASH. b, Correlation comparisons between hepatic senescence markers and COL1A1 or αSMA mRNA levels. c, RT–qPCR analysis of hepatic BMP4 and GREM1 mRNA levels in lean individuals, patients with NAFLD and patients with NASH. d, RT–qPCR analysis of hepatic mRNA expression of BMP4 and GREM1 in lean (n = 12), obese (n = 24) and diabetic–obese (n = 22) individuals. e, Correlation between hepatic BMP4 and GREM1 mRNA levels. f, RT–qPCR analysis of hepatic ID1 and ID2 mRNA expression in lean individuals, patients with NAFLD and patients with NASH. g, RT–qPCR analysis of ID1, ID2 and ID3 mRNA levels in IHH cells including the effect of BMP4, in the presence or absence of GREM1, on expression of these genes (n = 4 biologically independent experiments). h, Correlation comparison between mRNA levels of hepatic GREM1 and senescence markers. Data were collected in lean participants (n = 11–12), patients with NAFLD (n = 17–22) and patients with NASH (n = 19–24). Associations were determined using Spearman correlation analysis. Values are mean ± s.e.m. Statistical significance was determined by one-way ANOVA with post hoc Tukey’s test or Kruskal–Wallis with post hoc Dunn’s test.Source data

Fig 4: TGF-ß-mediated tenogenic differentiation(A) optohiMSCs were plated in monolayers on black-walled 24-well plates and allowed to reach 70%–80% confluency over 2 days. They were then switched to basal tenogenic medium (TM) with no TGF-ß (-T), 10 ng/mL TGF-ß3 (+T), or 3 or 6 µW 488 nm light with 5 min pulses every 2 h (+L3, +L6) for 14 days total.(B) qRT-PCR for collagen I (COL1A1) and scleraxis (SCX).(C) Immunofluorescent staining for COL1A1 (green), DAPI (blue), and SCX (purple). Scale bar, 200 µm.All data represent means ± SD with n = 7 in (B). *p < 0.05, ****p < 0.0001 as calculated by one-way ANOVA and post hoc Tukey-Kramer’s multiple comparisons test.

Fig 5: STING inhibitor in combination with dexamethasone alleviates remodeling in steroid resistant COPD fibroblasts.Comparison of COL1A1 (A), COL3A1 (B), and TGF-β (C) gene expression upon stimulation for 4 h with LPS, then treatment with Dex, H151 and combination in smoker COPD fibroblasts. Data representative of two independent experiments from four unique donors. Representative western blot analysis and densitometric analysis of remodeling mediators in smoker COPD fibroblasts at indicated concentration of the stimulant and treatment for 24 h (D-I). β-actin was used as a loading control. Data representative from two independent experiments. Full blots are supplemented in Fig D in S1 Raw images. (J) The gene expression of YAP1 was compared upon stimulation for 4 h with LPS, then treatment with Dex, H151 and combination in smoker COPD fibroblasts. Data representative from four unique donors. Representative western blot (K), and densitometric analysis of active YAP1 in smoker COPD fibroblasts at indicated concentration of the stimulant and treatment for 8 h (L). β-actin was used as a loading control. Data representative from two independent experiments. Full blots are supplemented in Fig D in S1 Raw images. Results are presented as mean (± SEM) and relative to unstimulated control. The values were compared across the different groups using one-way ANOVA followed by the post hoc Bonferroni test for multiple comparisons. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 vs. LPS stimulation. ###p < 0.001, ####p < 0.0001 vs. unstimulated control.