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. 2008 Feb 25;180(4):803–812. doi: 10.1083/jcb.200708109

Figure 6.

Figure 6.

Localization of SAC1 to the Golgi down-regulates Golgi PI(4)P and slows anterograde traffic. (A) Quiescent NIH3T3 expressing flag-SAC1 or flag-SAC1-K2A was stimulated with 10% serum in the presence or absence of 10 μM SB203580. Pulse-chase analysis of protein secretion was conducted as described in Materials and methods. Data are means ± SD from three independent experiments. (B) COS7 cells infected with adenoviruses expressing either empty vector, flag-SAC1, flag-SAC1-K2A, or flag-SAC1-LZ were labeled with [3H]myo-inositol, and total cellular PIs were analyzed and quantified by HPLC analysis. Data are means ± SD from four to six independent experiments. *, P < 0.5 × 10−3. (C and D) Human fibroblasts infected with adenoviruses expressing either GFP-SAC1 (C, green) or GFP-SAC1-K2A (D, green) were grown in the presence of 15% (+serum) or 0.5% serum (quiescent) for 48 h. The cells were fixed, permeabilized, costained with anti-PI(4)P antibodies (red), and analyzed by immunofluorescence microcopy. Bars, 20 μm.