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. Author manuscript; available in PMC: 2012 Dec 15.
Published in final edited form as: Cell Host Microbe. 2011 Dec 15;10(6):563–576. doi: 10.1016/j.chom.2011.10.014

Figure 5. AAV2 Infection is GRAF1-Dependent.

Figure 5

(A) Transduction of HeLa and 293T cells expressing EGFP-tagged dominant negative GRAF1. Flow cytometry analysis was performed as described in Figure 1A and 4A.

(B) AAV2-luc and Ad5-luc transduction of 293T cells expressing GFP-GRAF1DN. Values are normalized to EGFP vector control.

(C) Binding and endocytosis of AAV2 in 293T cells expressing GFP-GRAF1DN. Values are normalized to EGFP vector control.

(D) AAV2-cherry and Ad5-cherry transduction of 293T cells following GRAF1 knockdown by siRNA. Numbers indicate mean red fluorescence of pictured low-magnification microscope fields. Similar results were obtained with HeLa cells (not shown). Bar, 100 μm.

(E) AAV2-luc and (F) Ad5-luc transduction of HeLa and 293T cells following GRAF1 knock-down by siRNA A or B. Values are normalized to scrambled siRNA controls (scr).

(G) AAV2-luc endocytosis in HeLa and 293T cells transfected with GRAF1 siRNA with or without dynasore treatment. Values are normalized to scrambled siRNA/DMSO controls.

(H) Confocal microscopy localization of GFP-GRAF1 (green) with AAV2 or Ad5 virions (red) in HeLa cells. Insets show higher magnifications of boxed areas. Values indicate the percentage of virus (red) overlapping with GRAF1 (green). Bar, 10 μm.

Error bars indicate SD, *p < 0.05, **p < 0.005.