Figure 2
Roles of Fak and Src in the suppression of anoikis in human IECs. A. HIEC (Undifferentiated; filled columns) and 30PC Caco-2/15 (Differentiated; open columns) adhering cell cultures were maintained 24 h serum-free (control) with PF573228 (Fak inhibitor) or PP2 (Src inhibitor), or maintained instead in suspension. CASP-3 relative activity was then established, by comparison to controls. Results obtained with -2PC Caco-2/15 cells were comparable to those shown here for HIEC cells. B. Representative (n ≥ 5) WB analyses of the activation of Src and Fak, and verifications of Fak-Src interactions, from -2PC (Undifferentiated) and 30PC (Differentiated) Caco-2/15 cell cultures maintained as in ( A ). Specific antibodies for pY576/577p125Fak, pY418p60Src and pY397p125Fak, as well as for respective total protein forms, were used. C-E. -2PC (Undifferentiated; filled columns) and 30PC (Differentiated; open columns) Caco-2/15 cells were maintained and processed as in ( A-B ), except that the relative pY576/577 levels of Fak ( C ), as well as the relative activation levels of Src ( D ) and Fak ( E ), were established in comparison to controls. B-E. Results obtained with HIEC cells were highly similar to those shown here for -2PC Caco-2/15 cells. A, C-E. Statistically significant (0.0001 ≤ P ≤ 0.001; n ≥ 5) differences between treated and control cultures are indicated by (*). Statistically significant (0.0005 ≤ P ≤ 0.005; n ≥ 5) differences between differentiated and undifferentiated IECs are indicated by (#).