Fig. 3

PAD2 depletion alters the morphology of MCF10DCIS.com cells. a Representative images of scrambled-shRNA and PADI2-shRNA cells in culture. Scale bar = 200 μm. The frames on the images to the left are enlarged on the middle and right images. The middle images show the morphology of multiple cells that are growing together; images on the right are the pictures of single cells. The black arrow indicates cell protrusion and membrane ruffling in the scrambled-shRNA cells. The white arrow shows the flattened appearance of the cytoplasmic region of PADI2-shRNA cells. b Total RNA was isolated from scrambled-shRNA and PADI2-shRNA cells. RHOA, RAC1, and CDC42 mRNA levels were determined by qRT-PCR (SYBR) using scrambled-shRNA as a reference and β-actin normalization. Data were analyzed using the 2 ^{-ΔΔ C(t)} method and are expressed as the mean ± SD from five independent biological replicates with three technical replicates for each biological replicate (* p < 0.05). c The whole cell lysates of MCF10DCIS.com scrambled-shRNA and PADI2-shRNA cells were immunoblotted with RhoA, Rac1, and Cdc42. The blot was also probed with anti-β-actin as a loading control