Skip to main content
. 2023 Feb 8;14:550. doi: 10.1038/s41467-023-36189-6

Fig. 1. Substrate stiffness induces Vgll3 expression and its nuclear translocation in myofibroblasts.

Fig. 1

a Schematic representation of the protocol for regulating myofibroblast differentiation using substrate stiffness. b Western blot detection of αSMA, periostin and GAPDH in Adherent (Ad), Non-adherent (Non), and Re-adherent (Re) cardiac myofibroblasts (n = 5 each). c mRNA levels of fibrosis-related genes in Ad, Non and Re cardiac myofibroblasts (n = 5 each). d Venn diagram of genes upregulated by substrate stiffness and in infarcted hearts. e mRNA levels of Vgll3 in Ad, Non, and Re cardiac myofibroblasts (n = 5 each). f–i Confocal immunofluorescence images of endogenous VGLL3 in myofibroblasts cultured in suspension or adherent culture (f), or plated on soft (1 kPa) and stiff (50 kPa) hydrogels (h). Graphs indicate the ratios of nuclear to cytoplasmic VGLL3 intensities in myofibroblasts cultured in adherent (Ad) or suspension (Su) culture (g), or plated on hydrogels with different elastic modulus (i). j Ratios of nuclear to cytoplasmic VGLL3 intensities in myofibroblasts attached for different durations on plastic plate. k Nuclear localisation of EGFP-VGLL3 in myofibroblasts (αSMA) in the left ventricle of MI murine hearts on day 3. White arrowheads in merged images indicated representative signals for VGLL3 in myofibroblasts. l Confocal immunofluorescence images of myofibroblasts treated with DMSO (0.5%), latrunculin A (Lat A, 2 μM), and Y27632 (80 μM). m–o Ratios of nuclear to cytoplasmic VGLL3 intensities in myofibroblasts treated with the cytoskeletal inhibitors (m), the integrin β1 inhibitor BTT-3033 (30 μM) (n), or FAK inhibitor VS-4718 (50 μM) (o). All experiments were performed at least three times. Data in (b, c, e, g, i, j), and (m–o) are shown as the mean ± SEM. P-values were determined using one-way ANOVA followed by Tukey’s range test in (b, c) (Col1a1 and Col1a2) and (e), Kruskal–Wallis followed by Dunn’s test in c (Acta2), (i, j, m), and two-sided Mann–Whitney’s U test in g, n, and o, *P < 0.05, **P < 0.01, ***P < 0.001. Scale bars in (f, h, k) and l = 20 μm. Source data are provided as a Source Data file.