Fig. 3
CPSF6 cooperates with MxB to inhibit the association of NUP358 with HIV-1 cores. a HeLa-Ctrl and HeLa-MxB cells were not transfected (Mock) or transfected with siRNA targeting CPSF6 (siCPSF6) or negative control (siNC). 48 h after transfection, the expression levels of CPSF6 and MxB were monitored by western blot. b siRNA depleted cells were incubated with HIV-1WT, infectivity was determined 48 h post infection. c siRNA depleted cells were incubated with HIV-1WT, qPCR analysis of HIV-1 Late RT DNA, 2-LTR circle DNA and integrated DNA was preformed 24 h post infection. Results were a summary of 3 independent experiments. d, e HeLa cells were first transfected with siRNA targeting CPSF6 (siCPSF6) or negative control (siNC), a second transfection was performed 24 h later with plasmid expressing MxB-HA protein or empty vector. Western blot for CPSF6 and MxB 96 h following siRNA transfection was shown (d). 72 h after the first siRNA transfection, cells were infected with HIV-1WT. Cells were fixed 6 h post infection and stained for HIV-1 capsid protein p24 (green), NUP358 (red) and DAPI (blue) for cell nuclei. Scale bar, 10 μm (e). f P24 spots, the percentage of the p24 signals detected in the nucleus and the percent p24 colocalizing with NUP358 in HeLa cells was counted in about 40 cells. Data were representative of 3 independent experiments. Statistical significance: NS: not significant, *p < 0.05, **p < 0.01 and ***p < 0.001