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. 2012 Sep 14;7(9):e45085. doi: 10.1371/journal.pone.0045085

Figure 1. Requirement for DIP in non-apoptotic membrane blebbing.

Figure 1

(A–F) Constitutively blebbing M2 melanoma cells were plated upon glass coverslips for 3 hrs, fixed, permeabilized and stained simultaneously with phalloidin and antibodies directed against mDia2 or DIP. Cells are visualized using a 63x objective with a confocal microscope. (G) DIP (pool or individual siRNAs) or GAPDH (control siRNA) depletion in M2 cells was confirmed 72 hrs post-transfection by i.p.-western blotting. Tubulin was used as a loading control. (H) Upon 72 hrs of siRNA-mediated DIP (pool or individual) or GAPDH depletion, blebbing M2 cells were quantified. The experiment was performed in triplicate with n>50 cells per condition. (I) M2 cells treated with GAPDH, DIP pooled or individual siRNAs for 72 hrs were plated upon glass coverslips, stained for phalloidin and were visualized using a 63x objective by confocal microscopy.