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Fig. 2. Dispersed TGN46 is a function of tGolgin-1 depletion. (A-F) HeLa cells transduced once with siRNA-1 (A,B), siRNA-2 (C,D) or siRNA-3 (E,F) were fixed 2 days later and analysed by IFM for endogenous tGolgin-1 (A,C,E) and TGN46 (B,D,F). Stars denote cells without tGolgin-1 labelling and arrowheads denote cells with significant residual levels of tGolgin-1. (G,H) HeLa cells were co-transduced with siRNA-1 and plasmids encoding either siRNA-1-resistant mouse tGolgin-1 (G-I) or irrelevant HA-epitope tagged Rab27a (J-L), then fixed 2 days later and analysed by IFM using antibodies specific for human tGolgin-1 (G,J), for both human and mouse tGolgin-1 (H,K), and for TGN46 (I,L), and appropriate fluorochrome-conjugated secondary antibodies. Arrows (G-I) denote cells with negligible labelling for endogenous human tGolgin-1 but that are reconstituted with mouse tGolgin-1; notice the ribbon-like labelling of TGN46. Arrowheads (J-L) denote cells with negligible labelling for endogenous human tGolgin-1; notice the dispersed TGN46 labelling in most of these cells.

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