Effect of LCP1 I232F overexpression on actin polymerization. (A) 32D cells expressing LCP1 I232F with dysplastic features were dual fluorescently labeled with anti-LCP1 antibody (red color) and an F-actin probe, phalloidin (green color). The labeled cells were imaged by confocal microscopy. The mutant-expressing cells exhibit increased level of F-actin and enrichment of F-actin and LCP1 at the cell cortex. (B) Flow cytometric quantification of F-actin levels in 32D cells expressing either wtLCP1 or I232F mutant grown under indicated conditions. (C) HeLa cells expressing either wtLCP1 or I232F mutant were imaged by confocal microscopy after dual labeling with anti-LCP1 antibody and phalloidin. (D) Subcellular fractionation of 32D cells expressing LCP1 (wt and I232F) upon 3 days of IL-3 treatment. DAPI, 4′,6-diamidino-2-phenylindole; H2AX, H2A histone family member X; MEK, mitogen activated protein kinase kinase; PARP1, poly (ADP-ribose) polymerase 1; VDAC, voltage-dependent anion channel.