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. 2009 Feb 26;28(8):1016–1028. doi: 10.1038/emboj.2009.47

Figure 1.

Figure 1

AKAP450 is present into a network associated to the cis-face of the GA. (A) RPE1 cells stained with A24 antibody. (B) NP-40-soluble (SF) and insoluble (IF) fractions from RPE1 cells were analysed by WB with A24 antibody. (C) RPE1 cells were transfected with scramble or knockdown siRNAs (AKAP-1 or AKAP-2), short hairpin expressing pSUPER (AKAP-1 shRNA) or lentiviral (AKAP-1 shRNA lentivirus) vectors. After 72 h, a total lysate was prepared and analysed by WB for AKAP450 or α-tubulin as a loading control. Alternatively cells were fixed and processed by IF with A24 antibody. D, depleted cells; ND, nondepleted cells. Arrowheads indicate the centrosome (CTR). (D–F) Double-labelled cells for AKAP450 (green) and GM130 (D), CTR433 (E) or Golgin245 (F) were analysed by confocal microscopy. Merged images are shown. Arrowheads point to the centrosome. Bars, 10 μm. Fluorescence intensity profiles (at bottom) correspond to lines drawn in images D, E and F (lines 1, 2 and 3 as indicated). (G) Quantification of colocalisation by ICA based on the Pearson's correlation coefficient. The values are means of 10 lines drawn at random positions in 16 different cells, each line contained 500 pixels in average. Errors bars indicate ±s.e.m.