Subcellular localization of selected P-body-interacting proteins. Immunofluorescence microscopy of (A) endogenous Rck/p54, hnRNP K, Dom3Z, and ATG12; (B) ectopically expressed USP4; or (C) ectopically expressed RO52 in proliferating U2OS cells. U2OS cells were transfected with a plasmid encoding GFP-Dcp1a to mark P-bodies (green foci) in panel A. Endogenous proteins were visualized using the cognate rabbit antibodies followed by secondary goat anti-rabbit conjugated to Alexa 555 (red staining). Flag-tagged USP4 or RH-tagged RO52 was visualized using the corresponding anti-epitope monoclonal antibody followed by secondary goat anti-mouse conjugated to Alexa 488 (green staining). P-bodies were visualized using rabbit anti-Rck/p54 antibody followed by secondary goat anti-rabbit conjugated to Alexa 555 (red).
