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Figure 1


Fig. 1. siRNA knockdown of expression of AMFR. (A) NIH-3T3 cells untransfected (UT) or transfected with Dicer AMFR siRNA (d-siRNA AMFR) or control siRNA (ctl siRNA) were western blotted with the 3F3A antibody against AMFR and antibody against beta-actin, as indicated. (B) NIH-3T3 cells untransfected (UT) or transfected with d-siRNA AMFR or control siRNA were fixed and immunofluorescently labelled with the 3F3A mAb and antibody against mtHSP70. Images were acquired with the same confocal settings (using a wide-open pinhole), and the bottom panels show digitally zoomed images of the inset (white box, first row), with the merge presenting 3F3A labelling in red and mitochondrial mtHSP70 labelling in green. (C) 3F3A labelling intensity was quantified from confocal images acquired with an open pinhole (see top row, panel B) of NIH-3T3 cells untransfected (UT) or transfected with d-siRNA AMFR or control siRNA. Total intensity per field was divided by the number of cells in the field (excluding cells touching the edges), determined by Hoechst nuclear labelling (data not shown). **P<0.001. Bar, 50 µm (top row); 10 µm (zoom, bottom row).