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. 2014 Sep 15;289(44):30443–30458. doi: 10.1074/jbc.M114.608992

FIGURE 5.

FIGURE 5.

PKC-α-regulates surface ErbB2 translocation into a juxtanuclear compartment. A, live SKBR-3 cells on coverslips were stained with Alexa Fluor® 488-conjugated anti-ErbB2 antibody in cold, followed by treatment with DMSO (vehicle control) or PMA (100 nm) at 37 °C for 1 h. The cells were paraformaldehyde-fixed, permeabilized with saponin (0.2% in PBS), stained for PKC-α (red) and imaged under a confocal microscope. B, SKBR-3 cells were transfected with a non-targeting siRNA (control) or a PKC-α-specific siRNA (a smartpool or siRNA no. 2) for 48 h. Live cells were stained with Alexa Fluor® 488-conjugated anti-ErbB2 antibody in cold and treated with PMA (100 nm) for 1 h at 37 °C. The cells were stained for PKC-α (red) and imaged as in A. C, SKBR-3 cells were transfected with a non-targeting siRNA (control) or a PKC-α-specific siRNA smartpool for 48 h as in B, and 40-μg aliquots of Triton X-100 lysate protein subjected to Western blotting with anti-PKC-α and β-actin (loading control) antibodies.