Fig. 2. Overexpression of C312 disrupts the localization of TGN resident proteins. HeLa cells (a-f,m-o) or stable transfectants of TRVb-1 cells expressing TGG (g-i) or TTF (j-l) were transiently transfected with C312 or C312(Y2187A), as indicated, and analyzed by IFM using antibodies to the indicated proteins and to the HA tag (' columns) and appropriate secondary RRX- and FITC-conjugated secondary antibodies. The cells in d-f were cotransfected with furin-HA (50-100 ng/six-well dish) and T7-epitope-tagged C312 (5-7 µg/six-well dish); anti-HA was used to detect furin and anti-T7 to detect C312. (a,d,g,j,m) Cells with low C312 expression (level I); (b,e,h,k,n) cells with high expression (level III);(c,f,i,l,o) selected cells in which semiquantitative analyses showed levels of HA staining comparable to those in b, e, h, k, and n.

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