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. 2012 Apr 25;287(26):21856–21865. doi: 10.1074/jbc.M112.348094

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Effects of DHHC3 or DHHC7 inhibition or silencing on PI4KIIα localization and TGN integrity. A, DHHC RNAi effects on PI4KIIα localization. HeLa cells transfected with DHHC siRNA for 68 h and Myc-PI4KIIα for the last 20 h were labeled with anti-Myc, anti-DHHC3 or anti-DHHC7, and anti-TGN46 antibodies. Scale bars = 10 μm. Boxed areas were enlarged to show merged images of Myc-PI4KIIα and DHHCs. B, effects of PI4KIIα or DHHC3 RNAi on the TGN and cis-Golgi. Cells transfected with siRNA against DHHC or PI4KIIα for 48 h were labeled with either anti-TGN46 (a TGN marker) or anti-GM130 (a cis-Golgi marker) antibody. Scale bars = 10 μm. C, effect of overexpression of monomer red fluorescent protein (mRFP)-tagged DN-DHHC3 or DN-DHHC7 on TGN integrity. Cells were stained with anti-TGN46 antibody. Cells were scored visually in a blinded fashion for compact (normal) versus expanded (abnormal) TGN46 staining. 100 cells were scored per condition. Scale bars = 10 μm. The percentage of monomer red fluorescent protein-positive cells with compact TGN46 staining is displayed in the bar graph. Ctrl, control.