Knockdown of Na+/H+ exchanger regulatory factor 1 (NHERF1) expression in breast cancer cells. (a) Breast cancer cell lines, as indicated, were analysed for NHERF1 expression by immunoblotting. Blots were stripped and re-probed with β-actin for loading control. (b) Short hairpin RNA (shRNA) sequences targeting two different positions of human NHERF1 mRNA, NHERF-786 and NHERF-910, were retrovirally transduced in T47D and MCF7 cells, as shown. Cells were also infected with empty virus (Babe) to be used as a control. Cells that expressed NHERF1 shRNA sequences were selected by 0.5 μg/ml of puromycin. NHERF1 expression in MCF7 and T47D stable cells and their parental lines was examined by immunoblotting. α-Tubulin blotting was used for normalisation.