Pab pml a301 167a

Lysates from transfected or infected cells were prepared in a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) loading buffer by boiling for 10 min at 95°C and sonication for 1 min. Proteins were separated on sodium dodecyl sulfate-containing 8 to 15% polyacrylamide gels and transferred to PVDF membranes (Biorad), followed by chemiluminescence detection using a FUSION FX7 imaging system (Vilber). Following antibodies were used: mAb FLAG M2 (Sigma-Aldrich), mAb Myc 9E10 (1-9E10.2; ATCC), mAb β-actin AC-15 (Sigma-Aldrich), mAb HCMV IE1 p63-27 [74 (link)], mAb RCMV IE1 (kindly provided by Sebastian Voigt), mAb PML 5E10 (kindly provided by Roel van Driel) was used to detect ratPML, and pAb PML A301-167A and A301-168A (Bethyl Laboratories) were used in combination to detect human PML.

Following antibodies were used to visualize PML-NB proteins: pAb-PML A301-167A and pAb-PML A301-168A (Bethyl Laboratories, Montgomery, TX, USA), mAb-PML G8 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), pAb-Sp100 GH3 for WB (kindly provided by H. Will, Hamburg, Germany), pAb-Sp100 B01 (Abnova, Taipeh, Taiwan), mAb-Daxx MCA2143 (Bio-Rad Laboratories, Hercules, CA, USA), mAb-Daxx E94 (Abcam, Cambridge, UK), pAb-ATRX H300 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), mAb-ATRX 39f (Sigma Aldrich, St. Louis, MO, USA). Following antibodies were used to stain for viral proteins: mAb-IE1 p63-27 [43 (link)], pAb-IE2 pHM178 [44 (link)], pAb-IE86 for simultaneous detection of IE1 and IE2 [45 (link)], mAb-UL44 [46 (link)], mAb UL69 [47 (link)], pAb-UL84 [44 (link)], mAb-MCP 28-4 [48 (link)], mAb-pp150 (a kind gift from the Sinzger lab, Ulm, Germany). β-Actin was stained using mAb-β-Actin AC-15 antibody (Sigma Aldrich, St. Louis, MO, USA).