Fig. 4.

The effect of Rh-TG2 stimulation on NHLFs lysate and matrix protein levels. a Representative Western blots of TG2, FN and αSMA and c matrix TG2, FN and monomeric TGFβ1 in NHLFs treated with exogenous Rh-TG2 (1 µg/ml) and untreated NHLFs. e Representative Western blots of p-SMAD3 and t-SMAD3 in control NHLFs and NHLFs treated with 1 µg /ml Rh-TG2 for 2 or 6 h. g Graph showing the presence of active TGFβ in IPF fibroblasts treated with Rh-TG2 (1 µg/ml) via the Mink TGFβ reporter system as described in the “Materials and methods”. h Representative Western blots of TG2 and FN in NHLFs treated with an ERK inhibitor (10 µM), a TGFβ neutralising antibody (20 µg/ml) or an Alk5 inhibitor (10 µM) in the presence of Rh-TG2 (1 µg/ml). Graphs based on densitometry analysis of Western blots with controls normalised to 1.0 are shown in b–i and j expressed as a mean fold change ± SEM. GAPDH was used to ensure equal loading. *, p < 0.05; **, p < 0.01; and ***, p < 0.001