Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free

Immunoprecipitation: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody [NB100-97821]

Immunoprecipitation: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody [NB100-97821] - Detection of human JARID1B by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from 293T cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-JARID1B antibody NB100-97821 (lot NB100-97821-3) used for IP at 6 ug per reaction. JARID1B was also immunoprecipitated by a previous lot of this antibody (lot NB100-97821-2). For blotting immunoprecipitated Jarid1B, NB100-97821 was used at 1 ug/ml. Detection: Chemiluminescence with an exposure time of 75 seconds.

Western Blot: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 regulates the ubiquitination of JARID1B through TRAF6(A) Immunofluorescence images show a co-localization of endogenous JARID1B and TRAF6 in PC3 cells. Scale bars represent 10 μm. (B) and (C) Co-immunoprecipitation analysis shows that endogenous TRAF6 physically interacts with JARID1B in PC3 cells, as shown by reciprocal co-immunoprecipitation between the two proteins (Also see Supplementary Figure S5). (D)In vivo ubiquitination assay shows that TRAF6 increases K63-linked ubiquitination of JARID1B and SKP2 inhibits TRAF6-mediated JARID1B ubiquitination. Cells were transfected with Flag-JARID1B, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 constructs as indicated. WCL indicates the whole cell lysates. (E) TRAF6 mediates JARID1B ubiquitination through lysine residue 242. HEK293T cells were transfected with Flag-JARID1B WT or Flag-JARID1B-K242R, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 plasmids as indicated. In vivo ubiquitination assay was performed in a standard procedure. WCL indicates the whole cell lysates. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 regulates the ubiquitination of JARID1B through TRAF6(A) Immunofluorescence images show a co-localization of endogenous JARID1B and TRAF6 in PC3 cells. Scale bars represent 10 μm. (B) and (C) Co-immunoprecipitation analysis shows that endogenous TRAF6 physically interacts with JARID1B in PC3 cells, as shown by reciprocal co-immunoprecipitation between the two proteins (Also see Supplementary Figure S5). (D)In vivo ubiquitination assay shows that TRAF6 increases K63-linked ubiquitination of JARID1B and SKP2 inhibits TRAF6-mediated JARID1B ubiquitination. Cells were transfected with Flag-JARID1B, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 constructs as indicated. WCL indicates the whole cell lysates. (E) TRAF6 mediates JARID1B ubiquitination through lysine residue 242. HEK293T cells were transfected with Flag-JARID1B WT or Flag-JARID1B-K242R, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 plasmids as indicated. In vivo ubiquitination assay was performed in a standard procedure. WCL indicates the whole cell lysates. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 regulates the ubiquitination of JARID1B through TRAF6(A) Immunofluorescence images show a co-localization of endogenous JARID1B and TRAF6 in PC3 cells. Scale bars represent 10 μm. (B) and (C) Co-immunoprecipitation analysis shows that endogenous TRAF6 physically interacts with JARID1B in PC3 cells, as shown by reciprocal co-immunoprecipitation between the two proteins (Also see Supplementary Figure S5). (D)In vivo ubiquitination assay shows that TRAF6 increases K63-linked ubiquitination of JARID1B and SKP2 inhibits TRAF6-mediated JARID1B ubiquitination. Cells were transfected with Flag-JARID1B, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 constructs as indicated. WCL indicates the whole cell lysates. (E) TRAF6 mediates JARID1B ubiquitination through lysine residue 242. HEK293T cells were transfected with Flag-JARID1B WT or Flag-JARID1B-K242R, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 plasmids as indicated. In vivo ubiquitination assay was performed in a standard procedure. WCL indicates the whole cell lysates. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 regulates the ubiquitination of JARID1B through TRAF6(A) Immunofluorescence images show a co-localization of endogenous JARID1B and TRAF6 in PC3 cells. Scale bars represent 10 μm. (B) and (C) Co-immunoprecipitation analysis shows that endogenous TRAF6 physically interacts with JARID1B in PC3 cells, as shown by reciprocal co-immunoprecipitation between the two proteins (Also see Supplementary Figure S5). (D)In vivo ubiquitination assay shows that TRAF6 increases K63-linked ubiquitination of JARID1B and SKP2 inhibits TRAF6-mediated JARID1B ubiquitination. Cells were transfected with Flag-JARID1B, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 constructs as indicated. WCL indicates the whole cell lysates. (E) TRAF6 mediates JARID1B ubiquitination through lysine residue 242. HEK293T cells were transfected with Flag-JARID1B WT or Flag-JARID1B-K242R, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 plasmids as indicated. In vivo ubiquitination assay was performed in a standard procedure. WCL indicates the whole cell lysates. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

Enforced KDM5B expression facilitates melanocytic lineage-directed elimination by TMECG.a Quantitation of mRNA after 24 h, 48 h, 72 h and 7 days of Cpd1 treatment of MaMel63a cells as assessed by qPCR. Mean +/- SD. Shown is one representative example. b Regulation of differentiation, cytokinesis, and mitotic spindle assembly genes as detected by cDNA microarray analysis after KDM5B shRNA knockdown in WM3734 cells (n = 1). c, d Immunoblotting of melanocytic lineage and (de-)differentiation markers after 24 h of KDM5B induction in WM3734Tet3G-KDM5B cells (c) and after 72 h of Cpd1 treatment in MaMel63a cells (d). Shown are representative data (n = 2). e Anti-MITF immunostaining (upper panel) and Fontana-Masson staining (lower panels) of CM melanoma tumor grafts from Cpd1-treated vs. control mice. f MTT cell viability assay of WM3734 cells. Representative example is shown left (mean +/- SD, n = 2) and corresponding IC50 values on the right. TMECG was either concurrently given together with Cpd1 (“con") or added 3 days after Cpd1 pre-treatment (“pre"). Readout was performed after 72 h of TMECG treatment. g Persister-state-directed therapy model in vivo. Left: schematic representation of treatment dosing and timing in immunodeficient NMRI-(nu/nu)-nude mice. Right: tumor volumes of WM3734 xenografts (endpoint at day 30). TMECG was either concurrently given together with Cpd1 (“con") or added one week after Cpd1 pre-treatment (“pre"). Mean +/-SEM (6 mice in TMECG and Cpd1 control group, five mice in “con" and seven mice in “pre" group). Significance was determined by two-sided Mann–Whitney test. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35650266), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

Enforced KDM5B expression facilitates melanocytic lineage-directed elimination by TMECG.a Quantitation of mRNA after 24 h, 48 h, 72 h and 7 days of Cpd1 treatment of MaMel63a cells as assessed by qPCR. Mean +/- SD. Shown is one representative example. b Regulation of differentiation, cytokinesis, and mitotic spindle assembly genes as detected by cDNA microarray analysis after KDM5B shRNA knockdown in WM3734 cells (n = 1). c, d Immunoblotting of melanocytic lineage and (de-)differentiation markers after 24 h of KDM5B induction in WM3734Tet3G-KDM5B cells (c) and after 72 h of Cpd1 treatment in MaMel63a cells (d). Shown are representative data (n = 2). e Anti-MITF immunostaining (upper panel) and Fontana-Masson staining (lower panels) of CM melanoma tumor grafts from Cpd1-treated vs. control mice. f MTT cell viability assay of WM3734 cells. Representative example is shown left (mean +/- SD, n = 2) and corresponding IC50 values on the right. TMECG was either concurrently given together with Cpd1 (“con") or added 3 days after Cpd1 pre-treatment (“pre"). Readout was performed after 72 h of TMECG treatment. g Persister-state-directed therapy model in vivo. Left: schematic representation of treatment dosing and timing in immunodeficient NMRI-(nu/nu)-nude mice. Right: tumor volumes of WM3734 xenografts (endpoint at day 30). TMECG was either concurrently given together with Cpd1 (“con") or added one week after Cpd1 pre-treatment (“pre"). Mean +/-SEM (6 mice in TMECG and Cpd1 control group, five mice in “con" and seven mice in “pre" group). Significance was determined by two-sided Mann–Whitney test. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35650266), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunofluorescence: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 inactivation induces an accumulation of ubiquitinated JARID1B in nucleolus of cells in vitro and in vivo for cellular senescence(A) Immunofluorescence (IF) images show a co-localization of endogenous JARID1B and Fibrillarin (FBL) in nucleoli of PC3 cells upon SKP2 knockdown (Also see Supplementary Figure S6A). FBL indicates a nucleolar marker. Right panel: quantification of PC3 cells showing an increase of JARID1B localization in nucleolus. Error bars represent means +/- SD. (B) IF images show a co-localization of endogenous JARID1B and K63-Ub in nucleoli of PC3 cells upon SKP2 knockdown. (C) Western blotting assay shows an increase of beta-galactosidase ( beta-Gal) in PC3 cells upon SKP2 knockdown. (D) IF images show JARID1B in nucleolus as indicated by arrows and beta-Gal in cytoplasm in senescent cells upon SKP2 knockdown. (E) The co-localization of endogenous JARID1B and Fibrillarin (FBL) in nucleoli of prostate tissues in Ptenpc−/−;Trp53pc−/−;Skp2−/− mutant mice (Also see Supplementary Figure S6B). Scale bars represent 10 μm for panel A, B, D and E. (F) The positive staining of beta-galactosidase in prostate tissues of Ptenpc−/−;Trp53pc−/−;Skp2−/− mice but not in that of Ptenpc−/−;Trp53pc−/− mice. Scale bars represent 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunofluorescence: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 inactivation induces an accumulation of ubiquitinated JARID1B in nucleolus of cells in vitro and in vivo for cellular senescence(A) Immunofluorescence (IF) images show a co-localization of endogenous JARID1B and Fibrillarin (FBL) in nucleoli of PC3 cells upon SKP2 knockdown (Also see Supplementary Figure S6A). FBL indicates a nucleolar marker. Right panel: quantification of PC3 cells showing an increase of JARID1B localization in nucleolus. Error bars represent means +/- SD. (B) IF images show a co-localization of endogenous JARID1B and K63-Ub in nucleoli of PC3 cells upon SKP2 knockdown. (C) Western blotting assay shows an increase of beta-galactosidase ( beta-Gal) in PC3 cells upon SKP2 knockdown. (D) IF images show JARID1B in nucleolus as indicated by arrows and beta-Gal in cytoplasm in senescent cells upon SKP2 knockdown. (E) The co-localization of endogenous JARID1B and Fibrillarin (FBL) in nucleoli of prostate tissues in Ptenpc−/−;Trp53pc−/−;Skp2−/− mutant mice (Also see Supplementary Figure S6B). Scale bars represent 10 μm for panel A, B, D and E. (F) The positive staining of beta-galactosidase in prostate tissues of Ptenpc−/−;Trp53pc−/−;Skp2−/− mice but not in that of Ptenpc−/−;Trp53pc−/− mice. Scale bars represent 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunofluorescence: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 inactivation induces an accumulation of ubiquitinated JARID1B in nucleolus of cells in vitro and in vivo for cellular senescence(A) Immunofluorescence (IF) images show a co-localization of endogenous JARID1B and Fibrillarin (FBL) in nucleoli of PC3 cells upon SKP2 knockdown (Also see Supplementary Figure S6A). FBL indicates a nucleolar marker. Right panel: quantification of PC3 cells showing an increase of JARID1B localization in nucleolus. Error bars represent means +/- SD. (B) IF images show a co-localization of endogenous JARID1B and K63-Ub in nucleoli of PC3 cells upon SKP2 knockdown. (C) Western blotting assay shows an increase of beta-galactosidase ( beta-Gal) in PC3 cells upon SKP2 knockdown. (D) IF images show JARID1B in nucleolus as indicated by arrows and beta-Gal in cytoplasm in senescent cells upon SKP2 knockdown. (E) The co-localization of endogenous JARID1B and Fibrillarin (FBL) in nucleoli of prostate tissues in Ptenpc−/−;Trp53pc−/−;Skp2−/− mutant mice (Also see Supplementary Figure S6B). Scale bars represent 10 μm for panel A, B, D and E. (F) The positive staining of beta-galactosidase in prostate tissues of Ptenpc−/−;Trp53pc−/−;Skp2−/− mice but not in that of Ptenpc−/−;Trp53pc−/− mice. Scale bars represent 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunofluorescence: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 inactivation induces an accumulation of ubiquitinated JARID1B in nucleolus of cells in vitro and in vivo for cellular senescence(A) Immunofluorescence (IF) images show a co-localization of endogenous JARID1B and Fibrillarin (FBL) in nucleoli of PC3 cells upon SKP2 knockdown (Also see Supplementary Figure S6A). FBL indicates a nucleolar marker. Right panel: quantification of PC3 cells showing an increase of JARID1B localization in nucleolus. Error bars represent means +/- SD. (B) IF images show a co-localization of endogenous JARID1B and K63-Ub in nucleoli of PC3 cells upon SKP2 knockdown. (C) Western blotting assay shows an increase of beta-galactosidase ( beta-Gal) in PC3 cells upon SKP2 knockdown. (D) IF images show JARID1B in nucleolus as indicated by arrows and beta-Gal in cytoplasm in senescent cells upon SKP2 knockdown. (E) The co-localization of endogenous JARID1B and Fibrillarin (FBL) in nucleoli of prostate tissues in Ptenpc−/−;Trp53pc−/−;Skp2−/− mutant mice (Also see Supplementary Figure S6B). Scale bars represent 10 μm for panel A, B, D and E. (F) The positive staining of beta-galactosidase in prostate tissues of Ptenpc−/−;Trp53pc−/−;Skp2−/− mice but not in that of Ptenpc−/−;Trp53pc−/− mice. Scale bars represent 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunofluorescence: Lysine (K)-specific Demethylase 5B/KDM5B/JARID1B Antibody - BSA Free [NB100-97821] -

SKP2 regulates the ubiquitination of JARID1B through TRAF6(A) Immunofluorescence images show a co-localization of endogenous JARID1B and TRAF6 in PC3 cells. Scale bars represent 10 μm. (B) and (C) Co-immunoprecipitation analysis shows that endogenous TRAF6 physically interacts with JARID1B in PC3 cells, as shown by reciprocal co-immunoprecipitation between the two proteins (Also see Supplementary Figure S5). (D)In vivo ubiquitination assay shows that TRAF6 increases K63-linked ubiquitination of JARID1B and SKP2 inhibits TRAF6-mediated JARID1B ubiquitination. Cells were transfected with Flag-JARID1B, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 constructs as indicated. WCL indicates the whole cell lysates. (E) TRAF6 mediates JARID1B ubiquitination through lysine residue 242. HEK293T cells were transfected with Flag-JARID1B WT or Flag-JARID1B-K242R, HA-Ub-K63-only, Myc-TRAF6 and Myc-SKP2 plasmids as indicated. In vivo ubiquitination assay was performed in a standard procedure. WCL indicates the whole cell lysates. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25596733), licensed under a CC-BY license. Not internally tested by Novus Biologicals.