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Fig. 2. Reduction of Cripto-1 expression or inhibition of Cripto-1 signaling rescues Netrin-1 expression in mammary epithelial cells overexpressing Cripto-1. (A) Results from reverse transcriptase-PCR and Western blot experiments show reduction of Cr-1 expression in HC-11/Cr-1 cells transfected with a specific anti-Cr-1 siRNA. (B) Western blot analysis of lysates from Cripto-1 (Cr-1)-overexpressing HC-11 cells treated with anti-Cr-1 siRNA (Cr-1+siRNA) shows increase in expression of Netrin-1 and decrease in expression of Neogenin to levels comparable to those detected in wild-type (WT) HC-11 cells. Cr-1/cntrl, control HC-11/Cr-1 cells not treated with anti-Cr-1 siRNA. (C) Representative western blot analysis shows an increase in expression of Netrin-1 and a decrease in expression in Neogenin in HC-11/Cr-1 cells treated with the c-Src inhibitor (PP2) or with the PI-3K/Akt inhibitor (LY) compared to non-treated control HC-11/Cr-1 cells. (D) Histograms summarize results from densitometric analyses of western blots performed in the experiments described in C. Error bars indicate s.d. Treatment of HC-11/Cr-1 cells with a MAPK inhibitor (PD) had no significant effect on expression of Netrin-1 or Neogenin. (*P<0.05 compared to levels in the control group).
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