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. Author manuscript; available in PMC: 2019 Feb 11.
Published in final edited form as: Mol Cancer Res. 2018 May 14;16(8):1319–1331. doi: 10.1158/1541-7786.MCR-17-0589

Figure 2. Rab11-FIP5 regulates membrane expression of α6 integrin and cancer cell migration.

Figure 2.

DU145 cells were either untreated or transfected with non-targeting siRNA (siCon), siRNA against Rab11-FIP5 (siFIP5) or siRNA against α6 integrin (si α6). (A) Flow histograms of cell surface expression of laminin binding integrin subunits α6 or α3 and unrelated adhesion receptors CD44 and E-cadherin using immunolabelling of receptors in fixed, non-permeabilized untreated (solid line, shaded), siCon (solid line), siFIP5 (dashed line) and si α6 (dotted line) cells. (B) Relative mean peak fluorescence values of cell membrane expression of the receptors in untreated, siCon, siFIP5 and si α6 cells (**p<0.01, n=5). (C) Total cell lysate of untreated (DU145), siCon (siCon) and cells treated with two different siRNA against Rab11-FIP5 (siFIP5 #1, #2) immunoblotted for Rab11-FIP5 (FIP5), α6, α3, Rab11a and α-tubulin. (D) Untreated and DU145 cells treated with non-targeting siRNA (siControl), α6 integrin function blocking antibody (GOH3), siRNA against Rab11-FIP5 (siFIP5#1) and (siFIP5#2) and were allowed to migrate in a modified Boyden chamber with 8μm pores in response to laminin enriched HaCaT conditioned media for 6 hours. Percent untreated or treated cells migrated were counted in 50 fields of view per sample (n=3, each experiment in triplicates). Statistical significance assessed by student’s unpaired t-test (**p<0.01, ***p<0.001).