Cell adhesion properties and integrin expression of AGR-2-silenced PC3 cells.

<p><b>A.</b> Adhesion properties of PC3^control and PC3^AGR2sh cells were evaluated following growth in culture dishes coated with various ECM proteins. First, cells were seeded in duplicate onto the coated substrates and allowed to adhere. Next, unbound cells were washed away, and the adherent cells were fixed and stained. Finally, the stain was extracted and quantified colorimetrically. Significant reduction in cellular adhesion to fibronectin (***p<0.001), collagen I (**p<0.01), collagen IV (***p<0.001), laminin (*p<0.05) and fibrinogen (*p<0.05) were observed in case of PC3^AGR2sh cells when compared to PC3^control cells. BSA coated wells were used as the reagent control (p>0.1). Data presented here are mean ± SEM. <b>B</b>. Significant down-regulation of α4, α5, αV, β3 and β4 integrins was observed in PC3^AGR2sh cells as compared to PC3^control cells. No difference in β1 integrin levels was observed between the two cell types. Beta-actin was used as the loading control. Multiple protein bands present in the blots include integrin precursor and mature proteins as well as cleaved products is expected molecular mass according to manufacturer’s information (Cell Signaling Technology, Cat# 4749S).