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. 2020 Oct 21;16(10):e1008946. doi: 10.1371/journal.ppat.1008946

Fig 1. p120 binds to HPV16 and promotes virus infection.

Fig 1

A. Summary of L2 immunoprecipitation-mass spectrometry performed on samples of HeLa cells infected for 16 hrs or uninfected cell extract incubated with purified HPV16 PsV as described in Inoue et al., 2018. B. HeLa cells infected with or without WT HPV16.L2F (Luc) for 6 hrs were lysed and the resulting extract subjected to immunoprecipitation using a FLAG antibody. The precipitated material was analyzed by SDS-PAGE and immunoblotting using the indicated antibodies. Samples labelled input were not immunoprecipitated. See S1B Fig for IgG control. C. HeLa cells were infected with HPV16.L2F (Luc) or HPV16 L1 VLP (virus-like particles containing L1 only). 2.5 hpi, cells were lysed and the resulting extract subjected to immunoprecipitation using an antibody against p120. The precipitated material was subjected to SDS-PAGE and immunoblotting using antibodies recognizing the indicated proteins. Samples labelled input were not immunoprecipitated. See S1C Fig for IgG control. D. HeLa cells transfected with the indicated siRNA were infected with WT HPV16.L2F (GFP-S), with or without γ-secretase inhibitor XXI. 48 hpi, cells were lysed and the resulting extract subjected to SDS-PAGE and immunoblotting using the indicated antibodies. Graph shows data normalized against WT HPV16.L2F (GFP-S)-infected cells treated with scrambled siRNA, and further normalized against the actin level. Data represent the mean ± SD of at least three independent experiments. E. As in D, except the human SiHa cervical cancer cells were used. F. As in D, except the human SCC-47 oropharyngeal cancer cells were used. G. HeLa cells transfected with the indicated siRNA were infected with HPV5, HPV16, or HPV18 pseudovirus harboring a luciferase reporter plasmid, with or without γ-secretase inhibitor XXI. 48 hpi, luciferase activity was measured from the cell culture media. Graph shows luciferase activity normalized against scrambled siRNA-treated cells with and without HPV. Data represent the mean ± SD of at least three independent experiments. The immunoblot reveals the extent of p120 (or PS1) depletion.