Fig. 1

Depletion of PAD2 suppresses cell migration in MCF10DCIS.com cells. a Total RNA was isolated from MCF10DCIS.com cells infected with scrambled-shRNA and PADI2-shRNA lentiviruses. PADI2 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 three independent experiments (*p < 0.05). b The whole cell lysates of MCF10DCIS.com scrambled-shRNA and PADI2-shRNA cells were immunoblotted with anti-PAD2 antibody. Anti-β-actin was used as loading control. c Immunofluorescent analysis was performed on scrambled-shRNA and PADI2-shRNA cells probed with anti-PAD2 antibody, (d) anti-pan-citrulline antibody, and DAPI (nuclei) staining. Scale bars = (C) 50 μm and (D) 25 μm. e Relative average areas of wound closure from the wound healing assays were calculated using ImageJ and normalizing to scrambled-shRNA (**p < 0.05). f A wound healing assays were performed on MCF10DCIS.com scrambled-shRNA and PADI2-shRNA cells. The cells were fixed using 4% paraformaldehyde 24 h after striking the wound (24 h.). The cells were then visualized and imaged using light microscopy to determine the extent of the wound closure. The widths of the initial wounds are indicated by the lines. The cells were also fixed immediately after striking the wound (0 h.) to indicate the size of the initial wounds. The images on the right are the enlarged images of the boxed images