Fig 2: Impact of depletion of SUMO1 or SUMO2/3 on degradation of selected proteins during HSV-1 infection.HepaRG cells depleted of SUMO1 or SUMO2/3 simultaneously were prepared by lentiviral transduction (see Methods) then analyzed by western blotting for the abundance of SUMO2/3 (A) or SUMO1 (B) conjugates after infection with wt HSV-1 (MOI 10) for 5 or 10 h in comparison with control cells (shNEG). The same samples were analyzed for the efficiency of viral gene expression (C) and the abundance of NACC1 (D), ZBTB10 (E) and ZBTB38 (F).

Fig 3: Confirmation that selected proteins of interest are present in His-SUMO2 purified samples and change in abundance during HSV-1 infection.Total and purified protein fractions from mock (m) and wt HSV-1 (wt; MOI 10 12 h p.i.) infected HA-HisSUMO2 cells were analyzed by western blot, with uninfected HA-His only total protein and purified fractions included as controls. Mock infected and infected samples are indicated by m and wt respectively. Probable sumoylated species of PML, NACC1, ZBTB10, ZBTB38, ZBTB4 and MORC3 were detected in the uninfected purified samples (indicated by dashes on the right of each panel), and were reduced in abundance during HSV-1 infection. ZBTB7A exhibited apparently increased SUMO2-modification in the infected purified fraction. RanGAP1 was used as an unchanged control for SUMO2-modification following HSV-1 infection. The predicted unmodified molecular weight of each protein is indicated in each panel, and where clear the likely major unmodified form is marked by an asterisk. For PML, this refers to the major isoforms PML.I and PML.II.