Validation of expression of integrins by RT-PCR and Western blot analysis. Expression levels of integrin α5 (A) and α6 subunits (B) were validated by RT-PCR and Western blotting. RT-PCR was done using the integrin α5- and α6-specific primers. GAPDH was used as the loading control. Cell lysates were prepared and Western blotted with anti-integrin α5 or anti-integrin α6 antibody. Vinculin blotting was used for equal loading. Integrin α5 overexpression (C) and α6 knockdown (D) were also validated by RT-PCR and Western blot. Stable cell lines overexpressing integrin α5 and knockdown α6 were established. RNA and protein lysates were used to demonstrate the expression of the plasmids.