Interaction of NCOR/SMRT Repressor Complexes with Papillomavirus E8^E2C Proteins Inhibits Viral Replication
Fig 5
The HPV16 and 31 E8^E2C proteins recruit the NCoR/SMRT complex to viral replication foci.
HeLa cells were transfected with 300ng of E1 expression vectors (pCMV neo 3xFlag-31E1 or pSG 3xFlag-16E1), 30 ng of E2 expression vectors (pSX 31 myc-E2 or pSG 16 E2) and 30 ng of the expression vectors for E8^E2C or the E8^E2C KWK mt proteins (pSG 31 E8^E2C HA/pSG 31 E8^E2C KWK mt HA or pSG 16 E8^E2C HA/pSG 16 E8^E2C KWK mt HA). Cells were stained with the indicated primary antibodies and analyzed by immunofluorescence microscopy. DNA was stained with DAPI (blue). Signals for E8^E2C (or the E8^E2C KWK mt) and NCoR/SMRT components in replication foci from 4–7 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 Student´s t-test (**p<0.01; ***p<0.001).