Fig. 4.

(A) Comparative analyses of AAV2-mediated transduction efficiency in HeLa cells after in vitro phosphorylation of AAV2 capsids by EGFR-PTK. HeLa cells were infected by ssAAV2-RFP vectors, which were pre-incubated with ATP, EGFR-TPK, or both. Transgene expression was detected by fluorescence microscopy 48 hrs post-infection. Original magnification 100×. (B) Quantitative analyses of AAV2 transduction efficiency in HeLa cells. Images from five visual fields were analyzed quantitatively by ImageJ analysis software. Transgene expression was assessed as total area of red fluorescence (pixel²) per visual field (mean ± SD). Analysis of variance (ANOVA) was used to compare test results with the control and they were determined to be statistically significant. *P < 0.05 vs. ssAAV2-RFP. (C) Infection of HeLa cells with scAAV2-EGFP vectors pre-incubated with ATP, EGFR-TPK, or both. Transgene expression was detected by fluorescence microscopy at 48 hrs post-infection. Original magnification 100×. (D) Quantitative analyses of AAV2 transduction efficiency was assessed as described above, and were determined to be statistically significant. *P < 0.05 vs. scAAV2-EGFP. (E) Comparative analyses of AAV2-mediated transduction efficiency in HeLa cells after in vitro phosphorylation of AAV2 capsids by CKII. HeLa cells were infected by ssAAV2-RFP vectors, which were pre-incubated with ATP, CKII, or both. Transgene expression was detected by fluorescence microscopy 48 hrs post-infection. Original magnification 100×. (F) Quantitative analyses of AAV2 transduction efficiency was assessed as described above. (G) Infection of HeLa cells with scAAV2-EGFP vectors pre-incubated with ATP, CKII, or both. Transgene expression was detected by fluorescence microscopy 48 hrs post-infection. Original magnification 100×. (H) Quantitative analyses of AAV2 transduction efficiency was assessed as described above.