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. 2009 Jul 29;83(19):9863–9874. doi: 10.1128/JVI.00539-09

FIG. 3.

FIG. 3.

Downregulation of protein levels of ectopically expressed Akt, but not p53, by HPV-18 E6*. (A) HA-tagged Akt was overexpressed in 293 cells in the absence (−) and presence (+) of untagged constructs expressing wild-type HPV-18 E6 (WT 18E6), the HPV-18 E6 splice mutant (18E6 SM) which contains a splice donor site mutation, or HPV-18 E6* (18 E6*). Cell extracts were separated by PAGE and then transferred to nitrocellulose and probed with anti-HA antibodies to quantitate residual Akt. After blots were stripped, they were reprobed with anti-β-galactosidase (βgal) antibodies to visualize coexpressed β-galactosidase as a control for transfection and sample loading. (B) Human p53 was ectopically expressed in p53-null Saos-2 cells in the presence or absence of constructs expressing wild-type HPV-18 E6, HPV-18 E6 containing a splice donor site mutation, or HPV-18 E6*. After PAGE separation and transfer to nitrocellulose, residual p53 was visualized by probing with monoclonal antibody DO-1. Transfection efficiency and equal sample loading were controlled for as described above.