Fig. 1.

Generation of MDA-MB-231 breast cancer cells stably over-expressing EBP50 WT, or EBP50 phospho-deficient or phospho-mimetic mutants. MDA-MB-231 cells were transfected with pBK-CMV-HA-EBP50 WT, EBP50 phospho-deficient S279A/S301A mutant, phospho-mimetic S279D/S301D mutant expression plasmids or the pBK-CMV-HA vector respectively and screened using G418 sulfate (300 μg/ml). Then, the cells stably expressing these plasmids were named WT, AA, DD or Vector cells respectively. The HA-tag was detected for exogenous EBP50 protein expression. The 6–9 clones of each, WT, AA, DD and vector control cells were mixed respectively before usage. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a protein-loading control.