ibrd9  (MedChemExpress)

Journal: bioRxiv

Article Title: A modular toolbox for in cellulo screening of small molecule inhibitors targeting chromatin reader domains

doi: 10.1101/2025.09.06.674632

Figure Lengend Snippet: (A) Schematic showing how nuclei isolation & permeabilization followed by flow cytometry can measure the retention of proteins on chromatin in lieu of high background fluorescence. Nuclei can be harvested and permeabilized from whole cells, and then washed to remove the unbound or weakly bound fraction of the protein of interest. Since the weakly bound fraction of protein is removed, the fraction of protein remaining can be measured at a better signal-to-noise ratio via flow cytometry. (B) Flow cytometry analysis of nuclei harvested from BRD9_BRD.1x or WT cells. N3 gate shows the GFP signal being measured in nuclei, after iBRD9 or control treatments. Treatments in the WT cell line show a change in autofluorescence in the nuclei from the drug treatments. (C) Normalized flow cytometry data showing how Acyl-eCRs with 1x or 2x copies of the BRD9 bromodomain remain bound to chromatin, after iBRD9 treatments. iBRD9 treatments were performed at 1 μM concentration for 24 hours. The percentage represents the GFP signal in treated cells as a ratio of the signal observed in untreated samples of the same cell type, after normalizing for the autofluorescence of the drug treatment in wild-type cells.

Article Snippet: The CBP/p300 bromodomain inhibitor: GNE-049 (MedChemExpress, HY-108435), CBP/p300 PROTAC: dCBP-1 (MedChemExpress, HY-134582), BRD4 bromodomain inhibitor: (+)-JQ-1 (MedChemExpress, HY-13030), BRD4 PROTAC: ARV-825 (MedChemExpress, HY-16954), BRD9 bromodomain inhibitor: iBRD9 (MedChemExpress, HY-18975), and broad-spectrum bromodomain inhibitor: Bromosporine (MedChemExpress, HY-15815) were dissolved in DMSO and then diluted to 1μM in mESC media for 24-hour treatments, unless stated otherwise.

Techniques: Isolation, Flow Cytometry, Fluorescence, Control, Concentration Assay

Journal: Cell Reports Medicine

Article Title: BRD9 inhibition overcomes oncolytic virus therapy resistance in glioblastoma

doi: 10.1016/j.xcrm.2025.102258

Figure Lengend Snippet: Pharmaceutically targeting BRD9 enhances the antitumor effect of oHSV1 in vitro (A) Flow cytometry analysis of oHSV1-treated control or IBRD9-pretreated (1 μM, 24 h) CT2A Nectin1 and MGG4 cells subjected to PI/Annexin V staining for cell death analysis (PI+) ( n = 3). (B) Quantitative real-time PCR analysis of oHSV1 glycoprotein D levels in control or IBRD9-pretreated (1 μM, 24 h) CT2A Nectin1 and MGG4 cells treated with oHSV1. GAPDH/Gapdh transcript normalization ( n = 3). (C) Plaque formation assay of oHSV1-treated control or IBRD9-pretreated (1 μM, 24 h) CT2A Nectin1 and MGG4 cell culture medium. The virus titer was determined after 48 h ( n = 3). (D) Calreticulin (CRT) exposure analysis of control or IBRD9-pretreated (1 μM, 24 h) CT2A Nectin1 cells treated with oHSV1 ( n = 3). (E) Calreticulin (CRT) exposure analysis of control or IBRD9-pretreated (1 μM, 24 h) MGG4 cells treated with oHSV1 ( n = 3). (F) Extracellular ATP level analysis in control or IBRD9-pretreated (1 μM, 24 h) CT2A Nectin1 and MGG4 cells treated with oHSV1 ( n = 3). (G) Extracellular HMGB1 level analysis in control or IBRD9-pretreated (1 μM, 24 h) CT2A Nectin1 and MGG4 cells treated with oHSV1 ( n = 3). (H) In vitro co-culture proliferation experiments with OT-I CD8 + T cells and cDC1s generated from WT mice in the IBRD9- and oHSV1-treated CT2A Nectin1 -OVA-B2m −/− cells ( n = 3). (I) Schematic of human glioblastoma-derived organoid processing and verification of oHSV1-mediated killing by PI staining, 3D cell titer assays, and ICD marker analysis. (J) PI staining of IBRD9-pretreated (1 μM, 24 h) human glioblastoma-derived organoids treated with oHSV1. Scale bars, 300 μm. (K) 3D cell viability assay of IBRD9-pretreated (1 μM, 24 h) human glioblastoma-derived organoids treated with oHSV1 ( n = 3). (L) Extracellular ATP-level analysis in control or IBRD9-pretreated (1 μM, 24 h) human glioblastoma-derived organoids treated with oHSV1 ( n = 3). (M) Extracellular HMGB1-level analysis in control or IBRD9-pretreated (1 μM, 24 h) human glioblastoma-derived organoids treated with oHSV1 ( n = 3). (N) Schematic of human glioblastoma-derived tumor slice processing and verification of oHSV1 replication by anti-HSV1 staining and quantitative real-time PCR. (O) Representative immunohistochemistry images of HSV1 staining in IBRD9-pretreated or control human glioblastoma-derived tumor slices. Scale bars, 50 μm. (P) Analysis of oHSV1 replication in IBRD9-pretreated (2 μM, 24 h) or control human glioblastoma-derived tumor slices. After oHSV1 treatment, the distribution of oHSV1 in the sections was detected by anti-HSV1 immunohistochemistry ( n = 3). (Q) Quantitative real-time PCR analysis of oHSV1 glycoprotein D levels in control or IBRD9-pretreated (2 μM, 24 h) human glioblastoma-derived tumor sections treated with oHSV1. GAPDH transcript normalization ( n = 3). Data represent mean ± SD. Two-way ANOVA (A, D, E, F, G, H, K, L, and M), unpaired two-tailed Student’s t test (B, C, P, and Q). The diagrams (I and N) were created using BioRender. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant.

Article Snippet: IBRD9 , Selleck , Cat# S7835.

Techniques: In Vitro, Flow Cytometry, Control, Staining, Real-time Polymerase Chain Reaction, Plaque Formation Assay, Cell Culture, Virus, Co-Culture Assay, Generated, Derivative Assay, Marker, Viability Assay, Immunohistochemistry, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: BRD9 inhibition overcomes oncolytic virus therapy resistance in glioblastoma

doi: 10.1016/j.xcrm.2025.102258

Figure Lengend Snippet: Pharmaceutically targeting BRD9 enhances the antitumor effect of oHSV1 in vivo , and BRD9 expression is associated with poor clinical outcome in cancer patients treated with oHSV1 (A) Survival curve of IBRD9, oHSV1, and ICB (the anti-mouse PD-1 antibody and the anti-mouse CTLA4 antibody) combination therapy in CT2A Nectin1 tumor-bearing mice ( n = 5). (B) Survival curve of IBRD9, oHSV1, and ICB (the anti-mouse PD-1 antibody and the anti-mouse CTLA4 antibody) combination therapy in GL261 Nectin1 tumor-bearing mice ( n = 5). (C) Survival curve of long-term survivor mice and age-matched control mice challenged with CT2A Nectin1 cells ( n = 5). (D) Survival curve of long-term survivor mice and age-matched control mice challenged with GL261 Nectin1 cells ( n = 5). (E) Kaplan-Meier analysis showing the PFS of patients with glioblastoma treated with oHSV1 subdivided by the expression of BRD9 (high-expression patients: n = 7; low-expression patients: n = 6). HR, hazard ratio. (F) Analysis of BRD9 expression in liver cancer and pancreatic cancer biopsy sections from patients participating in an oHSV1 clinical trial. Representative immunohistochemistry images of BRD9 staining (oHSV1 response: SD, stable disease; oHSV1 nonresponse: PD, progressive disease). Scale bars, 50 μm. (G) BRD9-stained sections were quantified by H-score (SD: n = 14; PD: n = 13). Data represent mean ± SD. Unpaired two-tailed Student’s t test (G) and log rank test (A–E). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant.

Article Snippet: IBRD9 , Selleck , Cat# S7835.

Techniques: In Vivo, Expressing, Control, Immunohistochemistry, Staining, Two Tailed Test