Fig. 1

LG283 inhibits the fibrogenic activity of cultured human dermal fibroblasts stimulated with TGF-β1. Human fibroblasts were pretreated with DMSO or 4.5 μM of DMSO-diluted LG283 for 1 h, followed by stimulation with 10 ng/ml human rTGF-β1 for an additional 24 h. a, b After harvest, mRNA and protein expression of indicated molecules were evaluated by real-time RT-PCR and western blotting, respectively. Values were normalized to GAPDH levels and are shown as the mean of fold change compared to vehicle control ± SEM. Molecular weights of the paneled proteins; 130-kDa (COL1A2), 250-kDa (FN1), 50-kDa (α-SMA), 55-kDa (pSmad3), and 37-kDa (GAPDH). All values represent mean ± SEM; n = 5 each group; *p ≤ 0.05; **p ≤ 0.01. c Effects of LG283 on mRNA expression of transcription factors associated with the mesenchymal transition in human dermal fibroblasts. All values represent mean ± SEM; n = 5 each group; *p ≤ 0.05. d Human dermal fibroblasts were immunostained for phospho-Smad3 (p-Smad3, green). Nuclear counter-staining was performed with DAPI. Representative images are shown on the left (40-fold magnification)