FIG. 6.

Effects of simultaneous or individual depletion of PML and Sp100 on HSV-1 infection in nontransformed human hepatocytes. HepaRG cells transduced with lentiviruses expressing shRNAs targeting luciferase (HALL), PML (HALP1), or Sp100 (HALS2) were isolated. These cell lines were then transduced a second time with lentiviruses with a different selectable marker expressing the same shRNAs to give cell lines HALL/L, HALP1/L, HALS2/L, HALP1/S2, and HALS2/P1 (L, S2, and P1 indicate shRNAs Luci, shSp100-2, and shPML1, respectively). (A) Western blot analysis of PML, Sp100, and an actin loading control in HepaRG, HALL, HALP1, HALS2, HALP1/S2, and HALS2/P1 cells. The various isoforms of PML and Sp100 are marked. The filter was probed sequentially for the two proteins. (B and C) The relative probability of plaque formation by either wt (in1863) (B) or ICP0-null mutant (dl1403/ CMVlacZ) (C) virus in the complete set of cell lines was tested by staining for β-galactosidase activity 24 h after low-MOI infection. HepA, naïve HepaRG cells. The experimental details of MOIs and dilutions used were similar to those outlined in the legend to Fig. 1. The error bars represent SEM.