Figure 6

Knockdown of integrin-α2 in TβRIII-KD MSL cells reverses migratory and invasive TβRIII-KD phenotypes. A) qRT-PCR analysis for ITGA2 mRNA expression before and after stable knockdown of integrin-α2 (α2-KD) in SUM159 cells with TβRIII-KD; graph bars represent the mean of the replicates with SEM error bars (***P <0.0001 for a two-tailed Student’s t-test). B) Flow cytometry analysis of α2-KD; arrow pointing to the left shows a shift towards a decrease in protein amount of integrin-α2 in TβRIII-KD SUM159 cell lines after α2-KD. C) Transwell migration assay representing number of cells migrated through transwell upon α2-KD in TβRIII-KD SUM159 cell line (ns = not significant, ***P <0.0001 for a two-tailed Student’s t-test); bar graph represents a mean of three replicates with SEM error bars. D) Transwell migration assay representing number of cells migrated upon treatment of TβRIII-KD SUM159 cell line with anti-α2 blocking antibody (ns = not significant, *P = 0.011, ***P <0.0001 for a two-tailed Student’s t-test). E) Transwell invasion assays with inserts pre-coated with matrigel allowing for testing the number of cells that can invade upon α2-KD in TβRIII-KD SUM159 cells (ns = not significant, ***P <0.0001 for a two-tailed Student’s t-test). F) Immunoblot analysis for phospho-ERK using protein harvested from SUM159 cells with TβRIII-KD and TβRIII-KD /α2-KD. MSL, mesenchymal stem-like; SEM, standard error of the mean; TβRIII-KD, type III transforming growth factor-beta receptor knockdown.