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Fig. 8. Overexpression of MMP-1 following Cdc42 silencing is mediated through the ERK1/2 pathway. (A) Western-blot analysis of whole-cell lysates from starved HSFs 72 hours after transfection with calcium phosphate alone (CaP), 20 nM irrelevant siRNA (siScr) or 20 nM siRNA targeting RhoA (siRhoA), Rac1 (siRac1) or Cdc42 (siCdc42). Cell lysates were analysed by immunoblotting with specific antibodies to phospho-ERK1/2, phospho-p38 and total ERK1/2 (ERK1,2). (B) Representative western-blot analysis of whole-cell lysates 72 hours after transfection (cell lysates) and of serum-free medium conditioned for 16 hours between days 2 and 3 after transfection (C.M.). HSFs were transfected with 20 nM irrelevant siRNA (siScr) or with 20 nM siRNA targeting Cdc42 (siCdc42) and cultured between days 1 and 3 after transfection with the indicated concentrations of the MEK-kinase inhibitor U0126, of the PI3K inhibitor LY294 and of the p38-MAP-kinase inhibitor SB203580. Cell lysates and C.M. were analysed by immunoblotting with specific antibodies to Cdc42, ERK1/2 and MMP-1.
