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Fig. 11. The rescue of Cdc42 knock down represses MMP-1 overexpression. RT-PCR analysis of Cdc42 and mutated Cdc42 (mCdc42) mRNA, and 28S rRNA levels was performed with total RNA extracted from HS578T cells transfected with 20 nM irrelevant siRNA (siScr) or 20 nM first siRNA targeting Cdc42 (siCdc42) and 1 µg either empty pShuttle (vector) or pShuttle-mCdc42 (mCdc42). Total RNA was extracted 72 hours after transfection with the siRNA. (top left) The amplification of mCdc42 mRNA performed with total RNA extracted from HS578T cells 48 hours after transfection with pShuttle-mCdc42, with (+RT) or without (–RT) the reverse transcription step. Representative western-blot analysis with specific antibodies to Cdc42, ERK1/2 and MMP-1 of whole-cell lysates and of serum-free medium conditioned (C.M.) by HS578T cells transfected with 20 nM irrelevant siRNA (siScr) or 20 nM first siRNA targeting Cdc42 (siCdc42) and 1 µg empty pShuttle (vector) or pShuttle-mCdc42 (mCdc42). (bottom) Densitometric analysis of MMP-1 measurements by western-blot analysis of C.M. Results are the means±s.d. of three independent experiments.

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