HPV 31 E8^E2C recruits NCoR/SMRT complexes to distinct nuclear foci.

RTS3b were transfected with a HPV31 URR reporter plasmid and expression vectors for E1 (500 ng) (pCMV neo 3xFlag-31E1), E2 (50 ng) (pSX 31 myc-E2) and E8^E2C or the E8^E2C KWK (50 ng) (pSG 31 E8^E2C HA or pSG 31 E8^E2C KWK mt HA). Cells were stained with (A) α-flag to detect E1 and α-HA to detect E8^E2C (KWK mt) proteins or (B) α-TBLR1 and α-HA to detect E8^E2C (KWK mt) proteins. Slides were then analyzed by immunofluorescence microscopy. DNA was stained with DAPI (blue). Signals for E8^E2C (or the E8^E2C KWK mt) and TBLR1 in replication foci from 7–9 individual cells were analyzed for colocalization using optical sections and Zeiss Axiovision40 4.8.2 software. Colocalization is expressed as the Pearson´s correlation coefficient. Statistical significance between wt and mt was determined by Students t-test (***p<0.001).