anti brd4 rabbit polyclonal antibody  (Bethyl)

Journal: Scientific Reports

Article Title: Overexpression of JMJD6 drives immune evasion via the BRD4–IRF1–PD-L1 axis and promotes malignancy in gastric cancer

doi: 10.1038/s41598-025-30705-y

Figure Lengend Snippet: Graphical abstract. Graphical abstract illustrating the hypothetical mechanism by which JMJD6 promotes tumor progression and immune evasion in GC. JMJD6 is overexpressed in gastric cancer cells and promotes BRD4 expression, which upregulates IRF1 and consequently increases PD-L1 expression. Elevated PD-L1 expression on tumor cells inhibits T cell–mediated antitumor immunity, thereby facilitating immune escape.

Article Snippet: Anti-JMJD6 mouse monoclonal antibody (sc-28348; Santa Cruz Biotechnology, TX, USA), anti-PD-L1 rabbit monoclonal antibody (13684; Cell Signaling Technology, MA, USA), anti-ACTB rabbit monoclonal antibody (3700; Cell Signaling Technology), anti-BRD4 rabbit polyclonal antibody (A301-985A50; Bethyl Laboratories, TX, USA), and anti-IRF1 rabbit monoclonal antibody (8478; Cell Signaling Technology) were used.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Overexpression of JMJD6 drives immune evasion via the BRD4–IRF1–PD-L1 axis and promotes malignancy in gastric cancer

doi: 10.1038/s41598-025-30705-y

Figure Lengend Snippet: JMJD6 regulates BRD4, IRF1 and PD-L1 expression. ( a ) The knockdown of JMJD6 by transfection with siRNA-JMJD6 suppressed BRD4, IRF1 and PD-L1 in MKN74. In addition, the knockdown of BRD4 by transfection with siRNA-BRD4 suppressed IRF1 and PD-L1 in MKN74. In contrast, the knockdown of BRD4 did not suppress JMJD6 in MKN74. ( b ) Knockdown of JMJD6 suppressed PD-L1 and BRD4 expression in MKN74 gastric cancer (GC) cells. White dotted lines indicate nuclear boundaries. ( c ) Co-culture assay of GC cells and T cells. Under JMJD6 knockdown, T cells had more potent anti-tumor activity against GC cells compared with NC, and the proliferation ratio of GC cells was significantly decreased (mean ± SD, n = 3; error bars indicate SD, n = 3). ( d ) An impedance-based tumor-cell killing assay. The knockdown of JMJD6 increased the anti-tumor activity of T cells and inhibited the proliferation of GC cells. ( e ) JMJD6 overexpression using plasmid transfection promotes BRD4, IRF1 and PD-L1 expression. ( f ) A hypothetical model of the overexpression or activation of JMJD6 in GC cells.

Article Snippet: Anti-JMJD6 mouse monoclonal antibody (sc-28348; Santa Cruz Biotechnology, TX, USA), anti-PD-L1 rabbit monoclonal antibody (13684; Cell Signaling Technology, MA, USA), anti-ACTB rabbit monoclonal antibody (3700; Cell Signaling Technology), anti-BRD4 rabbit polyclonal antibody (A301-985A50; Bethyl Laboratories, TX, USA), and anti-IRF1 rabbit monoclonal antibody (8478; Cell Signaling Technology) were used.

Techniques: Expressing, Knockdown, Transfection, Co-culture Assay, Activity Assay, Over Expression, Plasmid Preparation, Activation Assay

Journal: Science Advances

Article Title: Tumor microenvironment–targeted PROTAC nanoparticle self-assembly broadly predicted by structural descriptors

doi: 10.1126/sciadv.adu2292

Figure Lengend Snippet: ( A ) Cell viability curves of NMC cells after treatment with free dBET6 or Fi-dBET6 NPs, as assessed by CellTiter-Glo. Dye-only control was treated at concentrations equivalent to that encapsulated into Fi-dBET6 NPs, as quantified by HPLC. N/A, not available. ( B ) Immunoblotting of BRD4 and β-actin in NMC cells after 24-hour treatment. ( C ) NanoBiT cells were treated with 0.01 μM of either free dBET6 or Fi-dBET6 NPs at t = 0. ( D ) NanoBiT cells were treated with free dBET6 or Fi-dBET6 NPs and washed three times with PBS, and the expression of BRD4 was monitored over 24 hours in the presence of Endurazine. Profiles are plotted as mean fractional relative luminescence units (RLU) values by normalizing to DMSO control. ( E ) Mean fluorescence intensity (MFI) of Fi-dBET6 NPs taken up by NMC cells following pretreatment with chlorpromazine (CPZ), a pharmacological inhibitor of endocytosis, as measured by flow cytometry. ( F ) Fi-dBET6 NP uptake and lysosomal colocalization 4 hours postwashout. ER, endoplasmic reticulum. Scale bars, 10 μm. The ROI (white box) is expanded in the second row of each condition. ( G ) Proposed mechanism of nanoPROTAC uptake and release. Data are means of technical replicates ± SEM [(A), (C), and (D)] where n = 3 or means of biological replicates ± SEM (E) where n = 3. Statistics were calculated using an ordinary one-way analysis of variance (ANOVA) with Dunnett’s post hoc test (E).

Article Snippet: Slides were then stained using a Discovery XT processor (Ventana Medical Systems-Roche) using primary antibodies against BRD4 (Bethyl, 50-156-1488), murine CD31 (Abcam, #ab182981), murine P-selectin (LSBio, #LSB3578) or human P-selectin (LSBio, #LSC78725).

Techniques: Control, Western Blot, Expressing, Fluorescence, Flow Cytometry

Journal: Science Advances

Article Title: Tumor microenvironment–targeted PROTAC nanoparticle self-assembly broadly predicted by structural descriptors

doi: 10.1126/sciadv.adu2292

Figure Lengend Snippet: ( A ) IF staining of P-selectin and CD31 in NMC tumor tissue. ( B ) Quantification of MFI (left) and representative fluorescence emission (right) of NP localization in tumors 24 hours postinjection of Fi-dBET6, Dex-dBET6 (untargeted control), free ICG, or free dBET6 (unlabeled). ( C ) Representative IHC of BRD4 in NMC tissue 48 hours posttreatment. ( D ) Quantification of BRD4 degradation in (C). ( E ) Pharmacokinetics of dBET6, as measured in plasma over time (data are means ± SEM where n = 4 biological replicates). ( F ) Nude mice engrafted subcutaneously with NMC cells were treated twice weekly with 15 mg/kg ip treatments of free dBET6, Fi-dBET6 NPs, vehicle, or untreated. ( G ) Tumor growth curves, ( H ) Kaplan-Meier survival curve, and ( I ) mouse weight change in NMC xenografts. Data are shown as individual biological replicates with means ± SEM, and statistics were calculated using one-way ANOVA with Dunnett’s post hoc test [(B) and (D)], multiple unpaired t tests with Holm-Sidak correction (E), unpaired t test of Fi-dBET6 versus free dBET6 (G), or Mantel-Cox survival analysis (H). ns, not significant; sc, subcutaneous.

Article Snippet: Slides were then stained using a Discovery XT processor (Ventana Medical Systems-Roche) using primary antibodies against BRD4 (Bethyl, 50-156-1488), murine CD31 (Abcam, #ab182981), murine P-selectin (LSBio, #LSB3578) or human P-selectin (LSBio, #LSC78725).

Techniques: Staining, Fluorescence, Control, Drug discovery, Clinical Proteomics