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Fig. 7. Targeting of Arl1(Q71L) to endosomes results in recruitment of p230, but not GCC88 or GCC185 to endosomal membranes. HeLa cells were transiently transfected with either (A) GFP-Arl1(WT) or (B) myc-SNX3-Arl1(Q71L), fixed and permeabilized. (A) Transfected cells were stained for p230 with human anti-p230 antibodies followed by Alexa 594 goat anti-human IgG. (B) Transfected cells were co-stained for myc-SNX3-Arl1(Q71L) and p230 and either GCC88 or GCC185, as indicated. myc-SNX3-Arl1(Q71L) was detected with mouse anti-myc monoclonal antibodies and Alexa 568 goat anti-mouse IgG, p230 detected with human anti-p230 antibodies and FITC-sheep anti-human IgG, and GCC88 and GCC185 detected with rabbit anti-GCC88 or anti-GCC185 antibodies followed by Cy5-labelled goat anti-rabbit IgG. Transfected cells expressing high levels of myc-SNX3-Arl1(Q71L) resulted in recruitment of endogenous p230 onto endosomal structures, whereas both GCC88 and GCC185 retained their normal Golgi localization. Bars, 10 µm.
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