JMJD2B Antibody

Western Blot: JMJD2B Antibody [NB100-74605]

Western Blot: JMJD2B Antibody [NB100-74605] - Detection of Human JMJD2B by Western Blot. Samples: Whole cell lysate (50 ug) from HeLa, 293T, and Jurkat cells. Antibodies: Affinity purified rabbit anti-JMJD2B antibody NB100-74605 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.

Immunoprecipitation: JMJD2B Antibody [NB100-74605]

Immunoprecipitation: JMJD2B Antibody [NB100-74605] - Detection of human JMJD2B by western blot of immunoprecipitates. Samples: Whole cell lysate (1 mg for IP; 20% of IP loaded) from HeLa cells. Antibodies: Affinity purified rabbit anti-JMJD2B antibody NB100-74605 (lot NB100-74605-2) used for IP at 6 ug/mg lysate. JMJD2B was also immunoprecipitated by a previous lot (lot NB100-74605-1) of this antibody For blotting immunoprecipitated JMJD2B, NB100-74605 was used at 1 ug/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.

Knockdown Validated: JMJD2B Antibody [NB100-74605]

Knockdown Validated: JMJD2B Antibody [NB100-74605] - Knockdown of JMJD2B reduces the intracellular TG level and represses PPARgamma2 expression in HepG2 cells. HepG2 cells in 6-well plates were transfected with scramble RNA or JMJD2B siRNA. The levels of JMJD2B, H3K9me, H3K9me2, and H3K9me3 were measured by western blotting. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S3A. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/s41598-018-31953-x), licensed under a CC-BY license.

Western Blot: JMJD2B Antibody [NB100-74605] -

Western Blot: JMJD2B Antibody [NB100-74605] - Gomisin N (GN) represses the expression of JMJD2B, PPAR gamma2, & PPAR gamma2 steatosis target genes in HepG2 cells. HepG2 cells were treated with GN (100 μM) for 12 h. (A) JMJD2B expression was assessed by qPCR & western blotting. The full-length western blots corresponding to truncated blots are given in Supplementary Figure S4. (B) PPAR gamma2 expression was measured by qPCR. (C,D) The expression of PPAR gamma2 steatosis target genes was assessed by qPCR. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: JMJD2B Antibody [NB100-74605] -

Western Blot: JMJD2B Antibody [NB100-74605] - Overexpression of JMJD2B increases intracellular TG levels & stimulates PPAR gamma2 expression in HepG2 cells. HepG2 cells in 6-well plates were infected with Ad-GFP or Ad-JMJD2B. (A) The levels of JMJD2B, H3K9me, H3K9me2, & H3K9me3 were measured by western blotting. The full-length western blots corresponding to truncated blots are provided in Supplementary Figure S2A. (B) Intracellular TG levels were measured by TG assay kit. (C) PPAR gamma2 expression was assessed by qPCR & western blotting. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S2B. (D,E) The expression of PPAR gamma2 steatosis target genes was evaluated by qPCR. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05, **p < 0.01 vs. Ad-GFP. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: JMJD2B Antibody [NB100-74605] -

Western Blot: JMJD2B Antibody [NB100-74605] - JMJD2B expression increases in hepatic steatotic cell & animal models. (A) HepG2 cells were incubated with a mixture of palmitic acid (PA) & oleic acid (OA) (1:2 ratio) at 800 μM concentrations for 24 h, & intracellular triglyceride (TG) levels were analyzed by a TG assay kit. JMJD2B mRNA & protein levels were examined by qPCR & western blotting, respectively. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S1A. (B) HepG2 cells were treated with T0901317 (10 μM) for 24 h, & intracellular triglyceride (TG) levels were measured by a TG assay kit. JMJD2B mRNA & protein levels were examined by qPCR & western blotting, respectively. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. The full-length western blots are presented in Supplementary Figure S1B. (C) Total RNAs were isolated from the livers of HFD-induced obese mice. The JMJD2B mRNA levels were assessed by qPCR. Data represent means ± SEM of 5 mice. *p < 0.05 vs. ND mice. ND: normal diet. HFD: high fat diet. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: JMJD2B Antibody [NB100-74605] -

Western Blot: JMJD2B Antibody [NB100-74605] - Knockdown of JMJD2B reduces the intracellular TG level & represses PPAR gamma2 expression in HepG2 cells. HepG2 cells in 6-well plates were transfected with scramble RNA or JMJD2B siRNA. (A) The levels of JMJD2B, H3K9me, H3K9me2, & H3K9me3 were measured by western blotting. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S3A. (B) Intracellular TG levels were measured by a TG assay kit. (C) PPAR gamma2 expression was assessed by qPCR & western blotting. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S3B. (D,E) The expression of PPAR gamma2 steatosis target genes was evaluated by qPCR. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. scramble RNA. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: JMJD2B Antibody [NB100-74605] -

Western Blot: JMJD2B Antibody [NB100-74605] - JMJD2B expression increases in hepatic steatotic cell & animal models. (A) HepG2 cells were incubated with a mixture of palmitic acid (PA) & oleic acid (OA) (1:2 ratio) at 800 μM concentrations for 24 h, & intracellular triglyceride (TG) levels were analyzed by a TG assay kit. JMJD2B mRNA & protein levels were examined by qPCR & western blotting, respectively. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S1A. (B) HepG2 cells were treated with T0901317 (10 μM) for 24 h, & intracellular triglyceride (TG) levels were measured by a TG assay kit. JMJD2B mRNA & protein levels were examined by qPCR & western blotting, respectively. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05 vs. no treatment. The full-length western blots are presented in Supplementary Figure S1B. (C) Total RNAs were isolated from the livers of HFD-induced obese mice. The JMJD2B mRNA levels were assessed by qPCR. Data represent means ± SEM of 5 mice. *p < 0.05 vs. ND mice. ND: normal diet. HFD: high fat diet. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: JMJD2B Antibody [NB100-74605] -

Western Blot: JMJD2B Antibody [NB100-74605] - Overexpression of JMJD2B increases intracellular TG levels & stimulates PPAR gamma2 expression in HepG2 cells. HepG2 cells in 6-well plates were infected with Ad-GFP or Ad-JMJD2B. (A) The levels of JMJD2B, H3K9me, H3K9me2, & H3K9me3 were measured by western blotting. The full-length western blots corresponding to truncated blots are provided in Supplementary Figure S2A. (B) Intracellular TG levels were measured by TG assay kit. (C) PPAR gamma2 expression was assessed by qPCR & western blotting. The full-length western blots corresponding to truncated blots are presented in Supplementary Figure S2B. (D,E) The expression of PPAR gamma2 steatosis target genes was evaluated by qPCR. Data represent means ± SEM of three independent experiments performed in triplicate. *p < 0.05, **p < 0.01 vs. Ad-GFP. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: JMJD2B Antibody [NB100-74605] -

Immunohistochemistry: JMJD2B Antibody [NB100-74605] - Adenovirus-mediated JMJD2B overexpression stimulated hepatic steatosis in vivo. C57BL/6 mice (8 weeks old) were injected with adenovirus Ad-GFP or Ad-JMJD2B. After injection, Ad-injected mice were fed a HFD for 2 weeks. (A) JMJD2B expression was determined by qPCR (B) Immunostaining of JMJD2B (scale bar = 50 μm). Representative photographs are shown. JMJD2B positive cells were counted in ten random areas at 400× magnification & analyzed by using image J. Quantification of immunostaining assay is represented as percentage of JMJD2B positive cells. (C) Hepatic triglyceride (TG) levels were measured by a TG assay kit. (D) Liver morphology, ORO & H&E staining (scale bar = 100 μm). Representative images are presented. (E) Serum total TG levels. (F) Serum total cholesterol levels. Data represent means ± SEM from 5 mice. *p < 0.05 vs. Ad-GFP-infected mice. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30214048), licensed under a CC-BY license. Not internally tested by Novus Biologicals.