Fig 3.

Dystonin localization in HFFF2 cells. (A) shControl or shDyst1 cells were fixed, and the distributions of dystonin (red) and alpha-tubulin (green) were determined using antibodies DST and GAR₅₆₈ and DM1A and GAM₄₈₈, respectively. Identical exposure times were used to collect images of shControl and shDyst1 cells. (B) shControl cells were infected with vUL35RFP1D1, a virus that encodes a VP26 capsid protein fused to the mRFP. Sixteen hours postinfection, cells were fixed and stained as described above, except that secondary antibodies were GAR₄₈₈ and GAM₆₄₇. Infected cells (i) were distinguished from noninfected cells (ni) through mRFP fluorescence (not shown). (C) HFFF2 cells were transfected with pGFP-hEB3, which encodes GFP fused to the plus-end-binding protein EB3 or with pEGFP-C1 as a control. Twenty-four hours after transfection, cells were fixed and immunostained for dystonin as described for panel A (red). GFP-EB3 and GFP are visualized through autofluorescence (green). An area from each cell image (dashed box) is enlarged. Scale bars, 20 μm.