Figure 4
AIF interacts with PDK4 and plays a role in EMT. (A) HEK293T cells were transfected with the pCMV6-Entry empty vector or pCMV6-FLAG-Myc-PDK4. Forty-eight hours post-transfection, the cells were lysed and FLAG IP was performed. Mass spectrometry was performed to analyze the immunoprecipitates, and the top five proteins identified in the immunoprecipitates are shown. (B) Immunoblotting was performed with samples prepared as in A to confirm the PDK4-AIF interaction. (C) A549 cells and HCC827 cells were transfected with siNTC pool no. 2, siAIF pool, or siPDK4 pool at 1 day and 3 days post-seeding, then lysed for immunoblotting. (D) HCC827 cells were transfected with siNTC pool no. 2, siAIF pool, and siPDK4 pool, treated with erlotinib (erl), and stained with crystal violet as in Figure 3C. (E) HCC827 cells were transfected as in D. Two days after the second transfection, the cells were stained with CM-H2DCFDA and ROS levels were analyzed using flow cytometry.