Fig. 5. .

MV stimulation of PC3 cells leads to increased β-actin deimination that is reduced upon PAD inhibition. PC3 cells were stimulated with BzATP to microvesiculate and total deiminated proteins immunoprecipitated using the F95 pan-deimination protein antibody. (a) Increased deiminated β-actin was observed in PC3 cells stimulated to microvesiculate with BzATP (Lane 3) compared to untreated, control PC3 cells (Lane 2). Upon treatment with 10 µM of the pan-PAD-inhibitor Cl-am prior to MV stimulation, a significant decrease was observed in the levels of deiminated β-actin (Lane 1). Equal amounts of protein extract immunoprecipitated with F95 were loaded as shown by Coomassie blue staining. (b) A bar chart representing deiminated β-actin band intensity as measured by protein densitometry and normalised intensity (calculated in Image J) from each of the treatments immunoblotted in (a).