Journal: Development (Cambridge, England)

Article Title: miR-126-5p promotes retinal endothelial cell survival through SetD5 regulation in neurons.

doi: 10.1242/dev.156232

Figure Lengend Snippet: Figure 8: SetD5 requires BRD2 to control Sema3A expression

Article Snippet: Complexes were immunoprecipated in the same conditions than for in vitro experiments using 1μg of SetD5 antibody or IgG control and BRD2 immunoblotting analysis were performed with Novus rabbit anti-BRD2 antibody (NBP1-30475).

Techniques: Control, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Histone demethylase KDM9 activates the CCND1/AKT pathway to promote the malignant progression of gliomas through interaction with BRD2

doi: 10.1007/s00018-025-05900-9

Figure Lengend Snippet: KDM9 regulates CCND1 transcription through interaction with BRD2. ( A ) Protein silver staining of IP via KDM9 antibody enrichment in LN229 cells. ( B ) Transcription factors in the protein mass spectrometry data are shown. Exogenous and endogenous IP experiments were performed to analyze the interaction between KDM9 and BRD4 ( C ) or BRD2 ( D ). ( E ) The IF assay was used to detect the expression and co-localization of KDM9 and BRD2 in LN229 cells. ( F ) After the transfection of pcDNA3.1-BRD2-His into LN229-shKDM9 cells for 48 h, ChIP-qPCR was performed to detect the enrichment of H4K20me2 at the CCND1 P4 promoter region (with IgG as negative control). ( G ) After the transfection of wild-type or mutant KDM9 plasmids into U251 cell lines for 48 h, luciferase reporter assays were conducted to detect CCND1 transcription activity (Left). After the transfection of pcDNA3.1-BRD2-His into LN229-shKDM9 cells for 48 h, luciferase reporter assays were performed to detect CCND1 transcription activity (Right). ( H ) After the transfection of shBRD2 plasmids into U251-KDM9-WT cell lines for 48 h, luciferase reporter assays were conducted to detect CCND1 transcription activity. ( I ) After the transfection of pcDNA3.1-BRD2-His into shKDM9 glioma cells for 48 h, a Western blot experiment was conducted to detect the protein expression of KDM9, BRD2, CCND1, AKT/P-AKT, mTOR/P-mTOR, and CD133. Data are shown as the mean ± SD. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: CD133 (18470–1-AP, 1:1000), Histone H4 (16047–1-AP, 1:500), CCND1 (60186–1-Ig, 1:2000), SMAD2 (12570–1-AP, 1:2000), P-AKT (66444–1-IG, 1:1000), AKT (60230–2-IG, 1:1000), Flag-Tag (66008–4-Ig, 1:5000), BRD2 (22236–1-AP, 1:1000), and BRD4 (28486–1-AP, 1:1000) were purchased from Proteintech (Chicago, IL).

Techniques: Silver Staining, Mass Spectrometry, Expressing, Transfection, ChIP-qPCR, Negative Control, Mutagenesis, Luciferase, Activity Assay, Western Blot

Journal: Frontiers in Oncology

Article Title: Targeting BET Proteins With a PROTAC Molecule Elicits Potent Anticancer Activity in HCC Cells

doi: 10.3389/fonc.2019.01471

Figure Lengend Snippet: BETd-260 induces degradation of BRD2, BRD3, and BRD4 in HCC cells. (A) HepG2 cell line was treated by BETd-260, HJB-97, or JQ1 as indicated for 24 h. The protein levels of BRD2, BRD3 and BRD4 were examined by western blot analysis. Actin was used as a loading control. (B) HepG2 cell line was treated by BETd-260 at 100 nmol/L for different times. The protein levels of BRD2, BRD3, and BRD4 were examined by western blot analysis. Actin was used as a loading control. (C) BEL-7402, SK-HEP-1, SMMC-7721, HuH-7, and MHCC97H cell lines were treated by BETd-260 at 100 nmol/L for 24 h. The protein levels of BRD2, BRD3 and BRD4 were examined by western blot analysis. Actin was used as a loading control. Data are representative of three independent experiments.

Article Snippet: The following antibodies were used for IHC: BRD2 (IHC-00612), BRD4 (HC-00396), BAD (A302-384A) from Bethyl Laboratories (Shanghai, China); BRD3 (ab264420) from Abcam (Shanghai, China); cleaved PARP (Asp214) (#32563), activated caspase-3 (#9664), and Ki-67 (8D5) (9449) from Cell Signaling Technology (CST, Shanghai, China), Anti-Mcl-1 (MAB828) from R&D Systems (R&D Systems, Shanghai, China).

Techniques: Western Blot, Control

Journal: Frontiers in Oncology

Article Title: Targeting BET Proteins With a PROTAC Molecule Elicits Potent Anticancer Activity in HCC Cells

doi: 10.3389/fonc.2019.01471

Figure Lengend Snippet: BETd-260 induces BET degradation, triggers apoptosis, and inhibits proliferation in HCC xenograft tissue in mice. BALB/c mice bearing HepG2 and BEL-7402 xenograft tumors were treated by a single intravenous dose of 5 mg/kg BETd-260 (BETd) for 24 h or vehicle (Veh). (A) Two to three mice were sacrificed and tumor tissue was harvested. The expression of BRD2, BRD3, and BRD4, Mcl-1, Bad, activated caspase-3, cleaved PARP, and Ki 67 was examined by immunohistochemistry staining. Representative photographs were presented. (B) The percentages of HCC tumor cells positively stained with BRD2, BRD3, and BRD4, Mcl-1, Bad, cleaved caspase-3 (c-Casp3), cleaved PARP (c-PARP), and Ki 67 were quantified under microscopy, and plotted. Data are representative of three independent experiments. ** p < 0.01.

Article Snippet: The following antibodies were used for IHC: BRD2 (IHC-00612), BRD4 (HC-00396), BAD (A302-384A) from Bethyl Laboratories (Shanghai, China); BRD3 (ab264420) from Abcam (Shanghai, China); cleaved PARP (Asp214) (#32563), activated caspase-3 (#9664), and Ki-67 (8D5) (9449) from Cell Signaling Technology (CST, Shanghai, China), Anti-Mcl-1 (MAB828) from R&D Systems (R&D Systems, Shanghai, China).

Techniques: Expressing, Immunohistochemistry, Staining, Microscopy