Fig. 7
Expression of hPML isoforms in MEFs restricts HIV-1 and SIVmac. a Immunofluorescence staining of hPML in PML-KO MEFs stably transduced with FLAG-tagged hPML isoforms I to VI. Images are representative of multiple observations. hPMLs were stained with an anti-FLAG antibody (green) and nuclear DNA was stained using Hoechst 33342 (blue). Scale bar 5 µm. b Effects of hPML isoforms on HIV-1 and SIVmac infectivity. PML-KO cells transduced with individual hPML isoforms were infected with increasing doses of HIV-1NL-GFP (left) or SIVmac-GFP (right). The percentage of infected cells was measured 2 days later by FACS. c Effects of hPML isoforms on HIV-1 and SIVmac LTR-driven GFP expression. GFP MFI values are shown for the experiments in b. d Effects of SAHA on hPML-dependent restriction of HIV-1 infectivity and LTR-driven GFP expression. MEF cells transduced with the individual hPML isoforms were infected with HIV-1NL-GFP at a CRFK MOI of 0.1. Ten days later, the cells were treated with either DMSO (no drug control) or SAHA for 48 h, followed by FACS. The percentage of infected cells (left panel) and GFP MFI (right panel) were assessed. The values represent the means of three independent experiments with standard deviations. The SAHA-dependent fold-increase in GFP MFI was compared between cells transduced with individual hPML isoforms and those transduced with the empty vector (Ctrl) using the two-tailed Student’s t test. The calculated P values are indicated on the graph. ns non-significant