Fig. 3. Parallel displacement of TGN46 and endogenous GRIP domain proteins at similar expression levels of C312. (a-j) HeLa cells that were transiently transfected with C312 were analyzed by three-color IFM using rat anti-HA, mouse anti-tGolgin-1 and sheep anti-TGN46 with AMCA-, RRX- and FITC-conjugated species-specific antibodies, respectively. (a-i) Representative images of staining patterns for HA-C312 (a,d,g), TGN46 (b,e,h) and tGolgin-1 (c,f,i) obtained with C312 expressed at level I (a-c), II (d-f) and III (g-i). (j) Semiquantitative analyses of total cell expression level of AMCA fluorescence (representing C312) in cells characterized as having a tight pericentriolar Golgi staining pattern (intact), diffuse paranuclear staining (diffuse), or diffuse cytoplasmic distribution (cytoplasmic) for TGN46 and endogenous tGolgin-1. C312 expression is plotted in arbitrary units on a log scale on the y axis; circles represent values for individual cells, and bars represent the median of all analyzed cells. (k,l) Cells transiently transfected with C312 were analyzed by IFM using antibodies to HA (k) and golgin-97 (l). (m,n) Cells transiently transfected with g97-C179 were analyzed by IFM using antibodies to HA (m) and to TGN46 (n).

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