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. 2016 Apr 11;12(4):e1005556. doi: 10.1371/journal.ppat.1005556

Fig 4. HPV1 and 8 E8^E2C proteins interact with NCoR/SMRT components in an E8-dependent manner.

Fig 4

(A) 293T cells or HeLa cells were transfected with expression vectors for HA-tagged HPV1 and 8 E8^E2C wt or K2A d3-10 mutant proteins and whole cell lysates were directly analyzed (input) or precipitated with α-HA antibody (IP) and subjected to immunoblot analysis with the indicated antibodies. (B) E8 consensus sequences for alpha-PV (a1, 5, 6, 7, 8, 9, 10, 11 and 13 subgroups) and beta-PV (b1-6 subgroups) were obtained by extracting and aligning HPV E8^E2C sequences using the papillomavirus episteme database (http://pave.niaid.nih.gov [35]) and WebLoGo version 2.8.2 [36]. (C) The conserved KLK motif is important for the interaction of HPV8 E8^E2C with TBLR1 and HDAC3 and the transcription repression function. HeLa or RTS3b cells were transfected with expression vectors for HPV8 E8^E2C-HA wt or KLK mt—HA proteins and whole cell lysates were directly analyzed (input) or precipitated with α-HA antibody (IP) and subjected to immunoblot analysis with the indicated antibodies. (D) HeLa (left graph) or RTS3b (right graph) cells were transfected with the empty vector or HPV8 E8^E2C or E8^E2C KLK mt expression vectors (1ng), pC18-Sp1-luc (100ng) and pCMV-gluc (0.3ng). Luciferase activities were determined as described in Fig 3. Data are derived from three independent experiments and error bars indicate the SEM.