Figure 4. Endogenous TRPC7 co-localizes with a marker for the cis-Golgi, mannosidase II, in a rat pituitary cell line, GH4C1.
(A) GH4C1 cells were fixed and processed for immunocytochemistry using a polyclonal rabbit anti-hTRPC7 antibody detected by a goat anti-rabbit Alexa Fluor® 488 (green) and a monoclonal anti-mannosidase II antibody detected by a goat anti-mouse Alexa Fluor® 594 (red). Overlap is depicted by yellow (merge). (B) Immunoblot demonstrating the specificity of the polyclonal rabbit antisera raised to TRPC7. Whole cell lysate from either untransfected (negative control) or hTRPC7-transfected HEK-293 cells (positive control) and GH4C1 cells were subjected to SDS/PAGE and then immunoblotted using the affinity-purified anti-TRPC7 serum. A band of approx. 100 kDa is detected in both the hTRPC7-transfected HEK-293 cells and the GH4C1 cells but is absent in the untransfected controls. Scale bar, 10 μm.