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Proteintech dcaf1 vprbp
Dcaf1 Vprbp, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/dcaf1+vprbp/VPRBP+Antibody/pmc12875578-202-169-171
Average 93 stars, based on 51 article reviews

dcaf1 vprbp - by Bioz Stars, 2026-09

93/100 stars

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Article Title: DNA damage induced by HIV-1 Vpr triggers epigenetic remodeling and transcriptional programs to enhance virus transcription and latency reactivation
Article Snippet: Cells were blocked using PBST supplemented with 5% bovine serum albumin (BSA, Fisher Bioreagents BP9703100), 10% goat serum (Sigma-Aldrich, G9023-10mL), and 0.3 M glycine for 2 hours at room temperature while rocking. .. After blocking, samples were incubated with primary antibodies against γH2A.X (1:300, Cell Signaling 9718), pCHK1 (1:50, Cell Signaling 2348), pCHK2 (1:200, Cell Signaling 2661), acetyl-histone H2AK5 (1:1,000, Cell Signaling 2576), acetyl-histone H2A.Z K4/K7 (1:1,000, Cell Signaling 75336), total H2B (1:1,000, Cell Signaling 12364), acetyl-histone H2BK5 (1:1,000, Cell Signaling 12799), acetyl-histone H2BK12 (1:1,000, Cell Signaling 5410), acetyl-histone H2BK15 (1:1,000, Cell Signaling 9083), acetyl-histone H2BK20 (1:1,000, Cell Signaling 34156), total H3 (1:1,000, Cell Signaling 4499), phospho-histone H3S10 (1:1,000, Cell Signaling 53348), acetyl-histone H3K9 (1:1,000, Cell Signaling 9649), tri-methyl-histone H3K9 (1:1,000, Cell Signaling 13969), acetyl-histone H3K14 (1:1,000, Cell Signaling 7627), acetyl-histone H3K18 (1:1,000, Cell Signaling 13998), acetyl-histone H3K27 (1:1,000, Cell Signaling 8173T), tri-methyl histone H3K27 (1:1,000, Cell Signaling 9733), total H4 (1:1,000, Cell Signaling 2935), acetyl-histone H4K5 (1:1,000, Cell Signaling 8647), acetyl-histone H4K8 (1:1,000, Cell Signaling 2594), acetyl-histone H4K12 (1:1,000, Cell Signaling 13944T), Rpb1 CTD (1:1,000, Cell Signaling 2629T), Phospho-Rpb1 (Ser2) (1:1,000, Cell Signaling 13499T), Phospho-Rpb1 (Ser5) (1:1,000, Cell Signaling 13523T), DNA-PKcs (1:200, Cell Signaling 12311S), DNA-PKcs (phospho S2056) (1:200, Abcam ab18192), DCAF1/VprBP (1:400, Proteintech 11612-1-AP), NF-κB p65 (1:300, Cell Signaling 82425S), c-Jun (1:400, Cell Signaling 9165T), Phospho-c-Jun (Ser63) (1:1,000, Cell Signaling 91952T), Phospho-c-Jun (Ser73) (1:800, Cell Signaling 3270T), Sp1 (1:500, Cell Signaling 9389T), Phospho-Sp1 (Ser101) (1:300, Active Motif 39758), or S9.6 DNA-RNA hybrid (1:200, Kerafast ENH001) in blocking buffer overnight at 4 °C. .. The next day, cells were washed 3× with PBS in 5-min intervals and then incubated with anti-mCherry conjugated to Alexa Fluor 594 (1:800, Invitrogen M11240 ), secondary anti-rabbit-IgG conjugated to Alexa Fluor 488 (1:800, Cell Signaling 4412), secondary anti-rabbit-IgG conjugated to Alexa Fluor 594 (1:800, Cell Signaling 8889) or secondary anti-mouse-IgG conjugated to Alexa Fluor 488 (1:800, Cell Signaling 4408) in blocking buffer at room temperature for 1 hour.

Incubation:

other:

Article Title: HIV-1 Vpr drives epigenetic remodeling to enhance virus transcription and latency reactivation
Article Snippet: After blocking, samples were incubated with primary antibodies against γH2A.X (1:300, Cell Signaling 9718), pCHK1 (1:50, Cell Signaling 2348), pCHK2 (1:200, Cell Signaling 2661), acetyl-histone H2AK5 (1:1000, Cell Signaling 2576), acetyl-histone H2A.Z K4/K7 (1:1000, Cell Signaling 75336), total H2B (1:1000, Cell Signaling 12364), acetyl-histone H2BK5 (1:1000, Cell Signaling 12799), acetyl-histone H2BK12 (1:1000, Cell Signaling 5410), acetyl-histone H2BK15 (1:1000, Cell Signaling 9083), acetyl-histone H2BK20 (1:1000, Cell Signaling 34156), total H3 (1:1000, Cell Signaling 4499), phospho-histone H3S10 (1:1000, Cell Signaling 53348), acetyl-histone H3K9 (1:1000, Cell Signaling 9649), tri-methyl-histone H3K9 (1:1000, Cell Signaling 13969), acetyl-histone H3K14 (1:1000, Cell Signaling 7627), acetyl-histone H3K18 (1:1000, Cell Signaling 13998 ) , acetyl-histone H3K27 (1:1000, Cell Signaling 8173T ) , tri-methyl histone H3K27 (1:1000, Cell Signaling 9733), total H4 (1:1000, Cell Signaling 2935), acetyl-histone H4K5 (1:1000, Cell Signaling 8647), acetyl-histone H4K8 (1:1000, Cell Signaling 2594), acetyl-histone H4K12 (1:1000, Cell Signaling 13944T), DCAF1/VprBP (1:400, Proteintech 11612-1-AP) or S9.6 DNA-RNA hybrid (1:200, Kerafast ENH001) in blocking buffer overnight at 4 degrees C. The next day, cells were washed 3x with PBS in 5-minute intervals and then incubated with anti-mCherry conjugated to Alexa Fluor 594 (1:800, Invitrogen M11240), secondary anti-rabbit-IgG conjugated to Alexa Fluor 488 (1:800, Cell Signaling 4412), secondary anti-rabbit-IgG conjugated to Alexa Fluor 594 (1:800, Cell Signaling 8889) or secondary anti-mouse-IgG conjugated to Alexa Fluor 488 (1:800, Cell Signaling 4408) in blocking buffer at room temperature for 1 h. After incubation, cells were washed 3x with PBS at 5-minute intervals and stained with NucBlue stain (Thermo Fisher, R37605) and imaged.

Image Search Results

(A) Electrostatic surface potential of a Vpr-DCAF1 co-complex. Surfaces associated with hyperactive or DDR-deficient Vpr mutants are colored in blue and red, respectively. (B, C) Immunofluorescence microscopy quantification of DDR activation following infection of HeLa or differentiated THP1 cells with Vpr WT or mutant viruses 48 hours post-infection ( n = 50 cells). Dashed boxes highlight hyperactive and DDR-deficient Vpr mutants associated with sub-surfaces colored blue and red, respectively, from . The data underlying this Figure can be found in . (D) Immunoblot analysis of histone marks in HeLa cells infected with indicated viruses 48 hours post-infection. The unmodified images underlying this Figure can be found in . (E) Circle graphs displaying amino acid variance at relevant positions identified in the mutagenesis screen with consensus residues in the middle of the circle. Sequences obtained from Los Alamos database ( n ≈ 4,100). (F) Immunofluorescence microscopy quantification of DDR activation and histone marks following infection of HeLa cells with Vpr WT , Vpr Y15R , or Vpr Y15H mutant viruses 48 hours post-infection ( n = 50 cells). Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001; ** p < 0.01; * p < 0.05. The data underlying this Figure can be found in .

Journal: PLOS Biology

Article Title: DNA damage induced by HIV-1 Vpr triggers epigenetic remodeling and transcriptional programs to enhance virus transcription and latency reactivation

doi: 10.1371/journal.pbio.3003621

Figure Lengend Snippet: (A) Electrostatic surface potential of a Vpr-DCAF1 co-complex. Surfaces associated with hyperactive or DDR-deficient Vpr mutants are colored in blue and red, respectively. (B, C) Immunofluorescence microscopy quantification of DDR activation following infection of HeLa or differentiated THP1 cells with Vpr WT or mutant viruses 48 hours post-infection ( n = 50 cells). Dashed boxes highlight hyperactive and DDR-deficient Vpr mutants associated with sub-surfaces colored blue and red, respectively, from . The data underlying this Figure can be found in . (D) Immunoblot analysis of histone marks in HeLa cells infected with indicated viruses 48 hours post-infection. The unmodified images underlying this Figure can be found in . (E) Circle graphs displaying amino acid variance at relevant positions identified in the mutagenesis screen with consensus residues in the middle of the circle. Sequences obtained from Los Alamos database ( n ≈ 4,100). (F) Immunofluorescence microscopy quantification of DDR activation and histone marks following infection of HeLa cells with Vpr WT , Vpr Y15R , or Vpr Y15H mutant viruses 48 hours post-infection ( n = 50 cells). Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001; ** p < 0.01; * p < 0.05. The data underlying this Figure can be found in .

Article Snippet: For coupling immunofluorescence microscopy with bimolecular fluorescence complementation, 48-hours post-transfection HeLa cells were fixed with 4% PFA, and immunofluorescence labeling was performed as described above (DCAF1, 1:400, Proteintech 11612-1-AP; GFP, 1:500, Cell Signaling 2956).

Techniques: Immunofluorescence, Microscopy, Activation Assay, Infection, Mutagenesis, Western Blot

(A) Immunofluorescence microscopy quantification of histone marks in HeLa and differentiated THP1 cells infected with indicated viruses 48 hours post-infection ( n = 50 cells). Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001. The data underlying this Figure can be found in . (B) Immunofluorescence microscopy quantification of histone marks following infection of HeLa cells expressing control or DCAF1 knockdown constructs 48 hours post-infection ( n = 50 cells). HeLa cells were sequentially transfected and infected with the indicated provirus and a shRNA construct that expresses a BFP reporter from a separate promoter. mCherry and BFP expressing cells were validated for loss of DCAF protein and changes to histone marks. Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001. The data underlying this Figure can be found in . (C) Representative fluorescence microscopy images and quantification of DCAF1 localization in HeLa, MDMs, and differentiated THP1 cells infected with the indicated viruses 48 hours post-infection ( n = 50 cells). Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001. The data underlying this Figure can be found in .

Journal: PLOS Biology

Article Title: DNA damage induced by HIV-1 Vpr triggers epigenetic remodeling and transcriptional programs to enhance virus transcription and latency reactivation

doi: 10.1371/journal.pbio.3003621

Figure Lengend Snippet: (A) Immunofluorescence microscopy quantification of histone marks in HeLa and differentiated THP1 cells infected with indicated viruses 48 hours post-infection ( n = 50 cells). Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001. The data underlying this Figure can be found in . (B) Immunofluorescence microscopy quantification of histone marks following infection of HeLa cells expressing control or DCAF1 knockdown constructs 48 hours post-infection ( n = 50 cells). HeLa cells were sequentially transfected and infected with the indicated provirus and a shRNA construct that expresses a BFP reporter from a separate promoter. mCherry and BFP expressing cells were validated for loss of DCAF protein and changes to histone marks. Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001. The data underlying this Figure can be found in . (C) Representative fluorescence microscopy images and quantification of DCAF1 localization in HeLa, MDMs, and differentiated THP1 cells infected with the indicated viruses 48 hours post-infection ( n = 50 cells). Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001. The data underlying this Figure can be found in .

Techniques: Immunofluorescence, Microscopy, Infection, Expressing, Control, Knockdown, Construct, Transfection, shRNA, Fluorescence

(A, B) Representative live cell fluorescence microscopy images of HeLa or differentiated THP1 cells expressing indicated constructs 48 hours post-transfection or -infection, respectively. For washout experiments, cells were either untreated or incubated with detergent for 15 min prior to live-cell imaging. (C) Live-cell fluorescence quantification of dual or singly fluorescence-positive cells from and ( n = 50 cells). The data underlying this Figure can be found in . (D) Diagrams of BiFC co-transfections and representative live cell fluorescence microscopy images of HeLa cells co-expressing indicated Vpr constructs. (E) Representative fluorescence microscopy images of Vpr (left) and DCAF1 (right) localization in HeLa cells infected with indicated viruses with quantification of DCAF1 cytoplasmic-to-nuclear ratio (Top) and induction of γH2A.X foci (Bottom) ( n = 45 cells). Cells were co-transfected with NTvenus-Vpr and CTvenus-Vpr constructs for 24 hours prior to live-cell imaging. Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001. The data underlying this Figure can be found in .

Journal: PLOS Biology

Article Title: DNA damage induced by HIV-1 Vpr triggers epigenetic remodeling and transcriptional programs to enhance virus transcription and latency reactivation

doi: 10.1371/journal.pbio.3003621

Figure Lengend Snippet: (A, B) Representative live cell fluorescence microscopy images of HeLa or differentiated THP1 cells expressing indicated constructs 48 hours post-transfection or -infection, respectively. For washout experiments, cells were either untreated or incubated with detergent for 15 min prior to live-cell imaging. (C) Live-cell fluorescence quantification of dual or singly fluorescence-positive cells from and ( n = 50 cells). The data underlying this Figure can be found in . (D) Diagrams of BiFC co-transfections and representative live cell fluorescence microscopy images of HeLa cells co-expressing indicated Vpr constructs. (E) Representative fluorescence microscopy images of Vpr (left) and DCAF1 (right) localization in HeLa cells infected with indicated viruses with quantification of DCAF1 cytoplasmic-to-nuclear ratio (Top) and induction of γH2A.X foci (Bottom) ( n = 45 cells). Cells were co-transfected with NTvenus-Vpr and CTvenus-Vpr constructs for 24 hours prior to live-cell imaging. Analyses performed using a one-way ANOVA; ns, not significant; *** p < 0.001. The data underlying this Figure can be found in .

Techniques: Fluorescence, Microscopy, Expressing, Construct, Transfection, Infection, Incubation, Live Cell Imaging

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