Figure 6

Differentiation state-distinct contributions of α2, α3, α5 and α6 integrin subunits, in the suppression of human IEC anoikis. A. Representative (n ≥ 3) double labeling-merged immunofluorescence micrographs of adhering HIEC cell cultures maintained 24 h serum-free (control) with mouse IgG’s, P1E6 (α2 binding activity-blocking mAb), P1D6 (α5 binding activity-blocking mAb), or GoH3 (α6 binding activity-blocking mAb). ISEL (green) was thereafter performed, with DAPI (blue) counter-staining of nuclei. B. Same as in ( A ), except that adhering 30PC Caco-2/15 cells were maintained 24 h serum-free (control) with mouse IgG’s, P1B5 (α3 binding activity-blocking mAb), P1D6 (α5 binding activity-blocking mAb), or GoH3 (α6 binding activity-blocking mAb). C. Adhering HIEC (Undifferentiated; filled columns) and 30PC Caco-2/15 (Differentiated; open columns) cell cultures were maintained 24 h serum-free (control) with mouse IgG’s, P1E6, P1B5, P1D6, or GoH3. ISEL assays were performed and compared to controls. Statistically significant (0.0001 ≤ P ≤ 0.001; n ≥ 3) differences between treated and control cultures are indicated by (*). Statistically significant (0.0005 ≤ P ≤ 0.005; n ≥ 3) differences between differentiated and undifferentiated IECs are indicated by (#). A-B. Original magnifications: 20X. A, C. Results obtained with -2PC Caco-2/15 cells were highly similar to those shown here for HIEC cells.