Fig. 1.

CPEB2A expression in various un-manipulated and manipulated cell lines. a COX-2, miR-655, miR-526b and CPEB2A were quantified in different (COX-2 disparate) breast cancer cell lines, relative to MCF7 cells. Cell lines (MDA-MB-231 and MCF7-COX-2) with high COX-2 and miRNAs expression showed low expression of CPEB2A (measured with an A/E probe). b Western blots for CPEB2 protein (about 60 kDa, presumably CPEB2 A, identified with an isoform nonselective antibody) revealed a similar trend, high expression of CPEB2 in poorly malignant, low COX-2 expressing MCF7 and T47D cells, and low expression in high COX-2 expressing and metastatic MDA-MB-231 and Hs578T cells. c, d CPEB2 knockout (KO) through a double nickase CRISPR plasmid in MCF10A cells resulted in downregulation of CPEB2 mRNA (shown with RT-PCR) and protein (shown with Western blot). CPEB2 protein expression was knocked out with 80% efficiency. e, f. SiRNA-mediated knockdown of CPEB2 mRNA using a pool of siRNAs in MCF7 cells shows 75% downregulation. Data presented as mean of 3 replicates ± SEM. (*) indicates p < 0.05, (**) p < 0.001, (***) p < 0.0002