Figure S7.
TOPORS knockout inhibits arsenic-induced PML degradation, and SIM2 is the major SUMO1-interacting element. Related to Fig. 9. (A) Immunoblots for TOPORS and tubulin from crude cell extracts of cells transfected with Cas9 and TOPORS #1 and TOPORS #2 guide RNA containing RNPs. (B) Representative microscopy images of U2OS YFP-PML-V WT cells transfected with Cas9 only or Cas9 with two different guides for TOPORS at t = 0 h and t = 16 h after 1 μM arsenic treatment. (C) Average PML body intensity over an 18-h time course of arsenic exposure in cells transfected with Cas9 only or Cas9-TOPORS RNPs. Solid lines represent average intensity normalized to t = 0 h, and shaded areas are SEMs (n = 10 fields of view). (D) Coomassie-stained whole gel images for the 4xSUMO1 and 4xSUMO2 ubiquitination assays using TOPORS and RNF4 SIM mutants as shown in Fig. 9 H. Source data are available for this figure: SourceData FS7.

TOPORS knockout inhibits arsenic-induced PML degradation, and SIM2 is the major SUMO1-interacting element. Related to Fig. 9. (A) Immunoblots for TOPORS and tubulin from crude cell extracts of cells transfected with Cas9 and TOPORS #1 and TOPORS #2 guide RNA containing RNPs. (B) Representative microscopy images of U2OS YFP-PML-V WT cells transfected with Cas9 only or Cas9 with two different guides for TOPORS at t = 0 h and t = 16 h after 1 μM arsenic treatment. (C) Average PML body intensity over an 18-h time course of arsenic exposure in cells transfected with Cas9 only or Cas9-TOPORS RNPs. Solid lines represent average intensity normalized to t = 0 h, and shaded areas are SEMs (n = 10 fields of view). (D) Coomassie-stained whole gel images for the 4xSUMO1 and 4xSUMO2 ubiquitination assays using TOPORS and RNF4 SIM mutants as shown in Fig. 9 H. Source data are available for this figure: SourceData FS7.

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