Figure S2. VCP/p97 interaction with PML... | Rockefeller University Press

Figure S2.

$VCP/p97 interaction with PML increases during arsenic treatment. (A) Coomassie stained SDS-PAGE gel fractionating anti-GFP affinity purifications from the indicated cell types with or without arsenic treatment (As). Gels were sliced into four sections per lane (as indicated) for in-gel tryptic digestion. Molecular weight markers are indicated in kD. (B) Principal component analysis of intensity data for 2,156 identified proteins. (C–E) Scatter plots showing fold change versus two-tailed student’s t test P value for the comparisons indicated on the x-axes (n = 4). FDR filtering values are indicated above each chart. (F) Venn diagram showing overlap between significant changers for parts C and D. The intersection is highlighted in Fig. 1 B. (G) Immunofluorescence analysis of PELP, MDN1, and FIB in U2OS PML−/− + YFP-PML using Alexa-594 secondary antibodies (left) and YFP fluorescence (center). Merged images with DAPI staining are shown (right panels). (H) In U2OS PML−/− + YFP-PML p97, RNF4 or SPRTN levels were ablated using siRNA or the SUMO and ubiquitin E1 enzymes inhibited with ML792 or TAK243, followed by 6 h treatment with arsenic. YFP-PML colocalization with p97 was measured by fluorescence and immunofluorescence. Source data are available for this figure: SourceData FS2.$

VCP/p97 interaction with PML increases during arsenic treatment. (A) Coomassie stained SDS-PAGE gel fractionating anti-GFP affinity purifications from the indicated cell types with or without arsenic treatment (As). Gels were sliced into four sections per lane (as indicated) for in-gel tryptic digestion. Molecular weight markers are indicated in kD. (B) Principal component analysis of intensity data for 2,156 identified proteins. (C–E) Scatter plots showing fold change versus two-tailed student’s t test P value for the comparisons indicated on the x-axes (n = 4). FDR filtering values are indicated above each chart. (F) Venn diagram showing overlap between significant changers for parts C and D. The intersection is highlighted in Fig. 1 B. (G) Immunofluorescence analysis of PELP, MDN1, and FIB in U2OS PML^−/− + YFP-PML using Alexa-594 secondary antibodies (left) and YFP fluorescence (center). Merged images with DAPI staining are shown (right panels). (H) In U2OS PML^−/− + YFP-PML p97, RNF4 or SPRTN levels were ablated using siRNA or the SUMO and ubiquitin E1 enzymes inhibited with ML792 or TAK243, followed by 6 h treatment with arsenic. YFP-PML colocalization with p97 was measured by fluorescence and immunofluorescence. Source data are available for this figure: SourceData FS2.

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