Fig. 4: β6-driven MMP13 upregulation is dependent on TGFβ signalling.

a GSEA plot showing enrichment of HALLMARK_TGFβ_signalling gene set (M5896) in β6 induced versus control 1089^iβ6 cells. Data are representative of two independent experiments. NES normalised enrichment score. b Expression of β6, phospho-, total SMAD2/3 and total SMAD4 upon doxycycline (1 µg/mL) treatment in 1089^iβ6 cells grown in 2D for 48 h. Blots are representative of two independent experiments. ci Immunofluorescence staining of DAPI (blue) and SMAD4 (pink) and (cii) quantification of nuclear SMAD4 intensity in 1089^iβ6 cells treated with treatment for 48 h, where each dot represents one cell. Scale bar = 100 µm (or 50 µm for inserts). ****p < 0.0001 (Two tailed T test). d Relative luminescence in 1089^iβ6 cells co-transfected with SMAD-SBE firefly and renilla firefly reporter constructs, treated with 48 h doxycycline (1 µg/mL), SB431542 (10 µM) and/or stimulated with TGFβ (5 ng/mL). e MMP13 mRNA expression in 1089^iβ6 cells treated doxycycline (1 µg/mL) and SB431542 (10 µM) for 48 h. *p < 0.05, **p < 0.01 (One-way ANOVA with multiple comparisons). Data are presented as mean ± SEM, with biological replicates indicated by different colours. Data are representative of three independent experiments.