Co-localization of PKCα with calnexin (A) or calreticulin
(B) in control COS-1 cells. Cells were processed for
co-immunofluorescence as described under “Experimental
Procedures.” For calnexin and PKCα co-localization, calnexin was
visualized with donkey anti-rabbit FITC conjugate, and PKCα was
visualized with donkey anti-mouse TRITC conjugate. Assembly of fluorescence
images (panels 1 and 2) indicates fusion-generated
yellow immunofluorescence (panel 4) outside the nuclei
(panel 5). For calreticulin and PKCα co-localization,
calreticulin was detected with donkey anti-rabbit FITC conjugate, and
PKCα was detected with donkey anti-mouse TRITC conjugate. Assembly of
fluorescence images (panels 1 and 2) indicates
fusion-generated yellow immunofluorescence (panel 4) outside
the nuclei (panel 5). Co-localization of PKCα or PKCδ
with UGT1A10His expressed in COS1 cells. Co-localization was determined by
immunofluorescence that probed UGT1A10 with donkey anti-rabbit FITC conjugate
and PKCα with donkey anti-mouse TRITC conjugate as described under
“Experimental Procedures.” C, assembly of fluorescence
images (panels 1 and 2) indicates fusion, which generated
yellow immunofluorescence (panel 4) outside the nuclei
(panel 5). D, co-localization of PKC δ and UGT1A10His
expressed in COS-1. Co-localization was analyzed by immunofluorescence with
PKCδ detected with donkey anti-rabbit FITC conjugate, and UGT1A10His was
detected with donkey anti-mouse TRITC as described under “Experimental
Procedures.” Assembly of fluorescence images (panels 1 and
2) merged to generate yellow fluorescence (panel 4)
outside the nuclei as shown (panel 5). Background immunofluorescence
images in non-transfected cells are shown in supplemental Fig. S4. Scale
bars represent 20 μm(A) or 10 μm(B–D).