Anti pml

Primary rabbit antibodies included anti-actin (Sigma-Aldrich; A5060), anti-Daxx (Upstate; 07-471), anti-PIAS1 (LsBio; LS-B9173), anti-PIAS2 (LsBio; LS-C108717), anti-PIAS3 (LsBio; LS-C98795), anti-PIAS4 (LsBio; LS-C108719), anti-PML (Bethyl Laboratories; A301-167A), and anti-Sp100 (SpGH [37 (link)]) antibodies. Primary mouse antibodies included anti-ICP0 (11060 [38 (link)]), anti-ICP4 (58S [39 (link)]), anti-Daxx (AbD Serotec; MCA2143), anti-SUMO1 (Invitrogen; 33-2400), anti-SUMO2/3 (Abcam; ab81371), anti-UL42 (Z1F11 [40 (link)]), and anti-VP5 (DM165 [41 (link)]) antibodies. Primary sheep antibodies included anti-SUMO1 (Enzo Life Sciences; BML-PW0505) and anti-SUMO2/3 (Enzo Life Sciences; BML-PW0510) antibodies. Secondary antibodies included peroxidase-conjugated anti-mouse IgG (Sigma-Aldrich; A4416), DyLight 680- or DyLight 800-conjugated goat anti-rabbit or -mouse IgG (Thermo), and Alexa Fluor 488-, 555-, or 633-conjugated donkey anti-rabbit, -sheep, or -mouse IgG antibodies (Invitrogen).

Antibodies used for the IF studies were as follows: rabbit polyclonal antibody (pAb) anti-PML (BETHYL; #A301-167A), mouse monoclonal antibody (mAb) anti-PML (Santa Cruz Biotechnology; #sc-966), rabbit pAb anti-TGN46 (Thermo Scientific; #PA5-23068), rabbit pAb anti-SUMO-1 (abcam; #ab11672), AlexaFluor (AF) 488-conjugated rabbit pAb anti-GFP (Molecular Probes; #A21311), mouse mAb anti-CD147 (Affinity BioReagents; #MA1-19202), mouse mAb anti-alpha-Tubulin (Cell Signaling; #3873S), AF647-conjugated phalloidin (Molecular Probes; #A22287), and goat AF-labeled secondary antibodies (Life Technologies; #A11034, #A21236). Rabbit polyclonal anti-Sp100 antibody was a kind gift from Dr. Hans Will (Heinrich Pette Institute, Hamburg, Germany) [59 (link)]. L1-specific mouse mAb 33L1-7 and rabbit pAb K75 were described previously [60 (link),61 (link)]. L2-specific mouse mAb 33L2-1 was also previously described [62 (link)]. Click-iT EdU Imaging Kit (Molecular Probes; #C10338) was used for Click-iT reactions to label EdU-labeled pseudogenomes. For western blot analysis, L1 was detected with mouse mAb HPV16 312F and L2 with mouse mAb 33L2-1, combined with peroxidase-conjugated AffiniPure pAb goat secondary anti-mouse (Jackson ImmunoResearch; #115-035-003).

The following antibodies were used for immunofluorescence or western blotting: Primary rabbit polyclonal: anti-actin (Sigma-Aldrich, A5060), anti-Daxx (Upstate, 07–471), anti-ATRX (Santa Cruz, H300), anti-PML (Bethyl Laboratories, A301-167A; Jena Biosciences, ABD-030), anti-Sp100 (GeneTex, GTX131569), anti-Mx1 (Santa Cruz, sc-50509; ProteinTech, 13750-1-AP), anti-ISG15 (ProteinTech, 15981-1-AP), anti-ISG54 (IFIT2, proteinTech, 12604-1-AP), and anti-histone H3 (abcam, ab1791). Primary mouse monoclonal: anti-HIRA (Millipore, 04–1488), anti-ICP0 (11060, [114 (link)]), anti-ICP4 (58s, [115 (link)]), anti-VP5 (DM165, [116 (link)]) anti-UL42 (Z1F11; [117 (link)]), and anti-PML (abcam, ab96051). Primary antibodies were detected using the following secondary antibodies: DyLight-680 or -800 conjugated goat anti-rabbit or -mouse (Thermo; 35568 and SA5-35571), Alexa -488, -555, or -647 conjugated donkey anti-rabbit or -mouse (Invitrogen; A21206, A21202, A31572, A31570, A31573, A31571), or HRP conjugated goat anti-mouse (Sigma-Aldrich, A4416).