Expression of Rab27a in HOG cell line. A. HOG cells cultured in GM were subjected to SDS–PAGE under non-reducing conditions and analyzed by immunoblotting with anti-Rab27a polyclonal antibody. Compared to positive controls, Mewo and HOM-2 cell lines, HOG cells show significant levels of Rab27a expression. B. RTqPCR quantification of relative Rab27a mRNA expression levels in HOG cells cultured in GM or DM. C. Immunoblot analysis of Rab27a expression in HOG cells cultured in GM or DM. HOG cells were subjected to SDS–PAGE under non-reducing conditions and analyzed by immunoblotting with anti-Rab27a polyclonal antibody Immunoblot assays showed a moderate increase of Rab27a in DM cultures. D. HOG cells cultured in GM or DM were fixed and processed for confocal immunofluorescence analysis with anti-Rab27a polyclonal antibody, detected using an Alexa Fluor 555 secondary antibody. The squares correspond to enlarged regions showing pericentrosomal localization of Rab27a, more scattered in the case of GM cultures. Images correspond to the projection of the planes obtained by confocal microscopy. (DIC: Differential Interference Contrast). All data are representative of, at least, 3 independent experiments. (a.u., arbitrary units).