Figure 1. PADI4 is expressed in human breast cancer cell lines.

(A) PADI4 mRNA expression normalized to GAPDH in human breast cancer cell lines as assessed by RT-qPCR. t-test for difference between basal and luminal subclasses. (B) PADI1-4 mRNA expression in representative breast cancer cell lines as assessed by RT-qPCR. Dunnett’s multiple comparison test with PADI4 as comparator. (C-D) Western blots of nuclear PADI4 protein expression in luminal (panel C) and basal A and basal B (panel D) breast cancer cell lines. Lamin B1, loading control. (E-F) Western blots of nuclear PADI4 following shRNA knockdown in MCF10Ca1h, MCF7E cells (E) and MDA-MB-231 LM2 cells (F) cells. (G) RT-qPCR of PADI4 following knockdown (mean +/−SD, n=3; t-test). (H) ELISA for total citrullinated histone H3. Results are normalized to shCON (mean +/− SD, n=3; Dunnett’s multiple comparison test with shCON as the comparator. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.