Fig. 2.

Expression of core activation genes in hepatic stellate cells (HSCs) following deactivation by basement membrane substrate (Matrigel™). Reverse transcription–quantitative polymerase chain reaction (RT–qPCR) for α-smooth muscle actin (SMA) (a) and tissue inhibitor of metalloproteinase (TIMP)-1 (b) was performed as described using RNA extracted from human HSC at the following time-points: quiescent HSCs immediately following isolation and prior to plating on tissue culture plastic (column A); HSCs cultured on tissue culture plastic for 24 h (column B); HSCs activated following reculture on tissue culture plastic for 7 days (column C) and activated HSCs recultured in Matrigel™ for 48 h with loss of activated phenotype (column D). Expression of α-SMA and TIMP-1 is shown normalized relative to housekeeping gene hypoxanthine–guanine phosphoribosyltransferase (HPRT). For (a), one-way analysis of variance (anova) with Bonferroni's multiple comparison test of A versus C, B versus C and C versus D found significant difference with a P-value of <0·0001. Other comparisons were not significant. For (b), all comparisons found significant difference with a P-value of <0·0001 except A versus D.