Fig. 2

A20 attenuates fibrotic responses elicited by transforming growth factor-ß (TGF-ß). Foreskin fibroblasts transiently transfected with A20 or empty vector were incubated with TGF-ß2 (10 ng/ml) for 24 h and harvested. RNA, whole cell lysates and media were isolated, and examined by qPCR (a) and western analysis (b). qPCR results, normalized with glyceraldehyde-3-phosphate dehydrogenase (GAPDH), are means ± SD of triplicate determinations (n = 3). Cgn I Type I collagen, L cell lysates, S secreted. Representative immunoblots (n = 3). Band intensities, normalized for tubulin in each lane are shown below. c Immunofluorescence confocal microscopy using antibody to αSMA (red). Nuclei identified by 6-diamidino-2-phenylindone (DAPI) (blue). Bar = 25 μm