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, Left panel: Histogram of cells 391 expressing the GFP fluorescence (green line: cells expressing stably the EGFP-AT1 392 receptors, black line: non-transfected HEK293 cells) Right panel: Histogram of cells 393 labelled with phycoerythrin (PE): intact cells were stained with anti-GFP antibodies and 394 secondary phycoerythrin-labeled antibodies (PE) Fluorescence intensities of non, p.395

, transfected HEK cells (black line) and cells expressing stably the EGFP-AT1 receptors 396

H. Antibodies, Anti-GFP antibody revealed bands only in cells expressing the EGFP- 406

, AT1 receptor whereas sc-1173 and sc-57036 anti-AT1 antibodies revealed unspecific 407 bands whether the receptor had been transfected or not, p.408

, B -Antibodies immunolabelling on EGFP-tagged AT1 cells. (a, d, g) Images collected 409

, in the green channel using epifluorescence microscopy showing the fluorescence of 410

, the EGFP-tagged receptor which is localized principally at the cell plasma membrane 411 and in some perinuclear compartments. (b, e, h) Images collected in the red channel, 412 stained with primary antibodies directed against GFP epitope in b, or AT1 receptor (e, 413 sc-1173, h, sc-57036) followed by a red secondary antibody

, colocalization with the GFP signal (displayed in the 415 channel overlay) is only detected with the anti-GFP labelling (b and c respectively), 416 whereas no plasma membrane signal (e and h) or colocalization (g and i) could be 417 detected when using anti-AT1 antibodies. Scale bar is 10 µM, p.418