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. Author manuscript; available in PMC: 2013 Oct 1.
Published in final edited form as: FEBS J. 2012 Sep 11;279(20):3925–3936. doi: 10.1111/j.1742-4658.2012.08752.x

Fig. 1.

Fig. 1

siRNA-mediated silencing of P-protein expression in HEK293T cells. HEK293T cells stably transected with doxycycline-inducible P2 RNAi were treated with 0.1µg/ml doxycycline for 96 h to knockdown expression of P2 protein. Whole cell lysates, purified ribosomes and cytoplasmic fractions were prepared post-treatment and separated by SDS-PAGE. Proteins were transferred to nitrocellulose, and probed with monoclonal antibody against the conserved C-termini of P-proteins (3BH5). P1 and P2 protein levels are expressed as a percentage relative to levels in undepleted cells. Levels were calculated from the band intensities, which were normalized to either P0 (whole cell lysates and purified ribosomes) or ß-actin (cytoplasmic fraction). Representative blots from 2–3 independent experiments are shown.