Western blot for EhC2A transfectants. A representative Western blot is shown with three biological replicates each for EhC2A (363–391), EhC2A (502–530), and EhC2A (363–391 scrambled control) shRNA transfectants. Results are representative of three biological replicates per shRNA transfectant with each sample run in triplicate. Each sample was also serially diluted 1:2, 1:4, and 1:8. Each membrane was probed anti-EhC2A and with anti-actin antibody as a loading control. The level of EhC2A protein in the scrambled control transfectant was defined as 100% (± 5%). The EhC2A (363–391) shRNA transfectant had strongly reduced levels of EhC2A protein: it was only 3.0 ± 0.4% of the scrambled control. The EhC2A (502–530) shRNA transfectant had no knockdown effect on EhC2A levels (106.1 ± 7.3%).
Linford et al. BMC Microbiology 2009 9:38 doi:10.1186/1471-2180-9-38