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Fig. 4. Co-localization of activated PKC-ß and RACK-I. To determine whether activated PKC-ß co-localizes with RACK-I, paired melanocyte cultures plated onto Labtek Chamber slides were treated with phorbol dibutyrate (PDBu) for 90 minutes to activate PKC. Control wells received DMSO only (A1 and B1). Then they were subjected to double immunostaining using polyclonal antibody against PKC-ß (A2) or PKC-{alpha} (B2) and monoclonal antibody against RACK-I (A3 and B3). Then cells were further incubated with FITC-conjugated secondary antibody against PKC-ß or PKC-{alpha} antibodies and rhodamine-conjugated secondary antibody against RACK-I antibody and analyzed as in Fig. 1. The images were overlaid to determine co-localization (indicated in yellow) between PKC-ß and RACK-I (A4) and PKC-{alpha} and RACK-I (B4).





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