Journal: Gene Therapy

Article Title: BRD9 promotes the progression of gallbladder cancer via CST1 upregulation and interaction with FOXP1 through the PI3K/AKT pathway and represents a therapeutic target

doi: 10.1038/s41434-024-00488-4

Figure Lengend Snippet: A Correlation of NOZ and GBC-SD cell proliferation with I-BRD9 dosage and time was detected by CCK-8 assay. B Expression of BRD9 protein was detected by western blot after I-BRD9 treatment of NOZ, GBC-SD cells. C Changes in the number of clones of NOZ, GBC-SD cells after I-BRD9 treatment probed by colony formation assay. D The effect of I-BRD9 on the proliferative capacity of NOZ and GBC-SD cells was detected by Edu method. E Subcutaneous tumors were resected in the I-BRD9 treatment group and the vehicle treatment group, respectively. F Removed subcutaneous tumors weight in vehicle or I-BRD9 treatment group. G Removed subcutaneous tumors volume of different weeks in vehicle or I-BRD9 treatment group. H No significant pathologic changes were seen in the heart, liver, spleen, or kidneys after I-BRD9 treatment.

Article Snippet: I-BRD9 was purchased from MedChemExpress, and 740Y-P (PI3K activator, #HY-P0175; 20 μM, treatment time 24 h) was purchased from MedChemExpress, and all the reagents were dissolved in DMSO.

Techniques: CCK-8 Assay, Expressing, Western Blot, Clone Assay, Colony Assay

Journal: Gene Therapy

Article Title: BRD9 promotes the progression of gallbladder cancer via CST1 upregulation and interaction with FOXP1 through the PI3K/AKT pathway and represents a therapeutic target

doi: 10.1038/s41434-024-00488-4

Figure Lengend Snippet: A Volcano plot showing differentially expressed genes identified after siRNA treatment. (Gray: genes with no statistically significant changes; blue: low expressed genes; red: overexpressed genes, p < 0.05). B RNA-seq data GO analysis and visualization of biological processes (BP), cellular components (CC) and molecular functions (MF). C KEGG pathway analysis of differential genes. D The effects of I-BRD9 and 740Y-P on the proliferative capacity of NOZ and GBC-SD cells were explored by colony formation experiments. E The proliferation of I-BRD9 and 740Y-P on NOZ and GBC-SD cells was detected by CCK8 assay to detect the proliferation of NOZ, GBC-SD cells by I-BRD9, 740Y-P. F Detection of PI3k pathway and CST1 expression by Western blot assay.

Article Snippet: I-BRD9 was purchased from MedChemExpress, and 740Y-P (PI3K activator, #HY-P0175; 20 μM, treatment time 24 h) was purchased from MedChemExpress, and all the reagents were dissolved in DMSO.

Techniques: RNA Sequencing, CCK-8 Assay, Expressing, Western Blot

Journal: Gene Therapy

Article Title: BRD9 promotes the progression of gallbladder cancer via CST1 upregulation and interaction with FOXP1 through the PI3K/AKT pathway and represents a therapeutic target

doi: 10.1038/s41434-024-00488-4

Figure Lengend Snippet: A qPCR and Western blot of CST1 under I-BRD9 treatment. B Detection of CST1 protein expression level by western blot. C CCK8 assay to detect the effects of I-BRD9 and CST1 OE on the proliferation of NOZ and GBC-SD cells. D Colony formation experiments to investigate the effects of I-BRD9 and CST1 OE on the number of clones of NOZ and GBC-SD cells. E Western blot assays for CST1, PI3k pathway expression. F Correlation of bromodomain-containing protein 9 (BRD9) and CST1 expression detected in gallbladder cancer specimens.

Article Snippet: I-BRD9 was purchased from MedChemExpress, and 740Y-P (PI3K activator, #HY-P0175; 20 μM, treatment time 24 h) was purchased from MedChemExpress, and all the reagents were dissolved in DMSO.

Techniques: Western Blot, Expressing, CCK-8 Assay, Clone Assay

Journal: Gene Therapy

Article Title: BRD9 promotes the progression of gallbladder cancer via CST1 upregulation and interaction with FOXP1 through the PI3K/AKT pathway and represents a therapeutic target

doi: 10.1038/s41434-024-00488-4

Figure Lengend Snippet: Model diagram of BRD9 regulation of the PI3K-AKT pathway in GBC. BRD9, as a key protein of the bromodomain protein family, positively regulates CST1 expression by promoting the transcription factor FOXP1. It further activates the PI3K-AKT pathway and participates in the growth and proliferation process of gallbladder cancer. And the selective small molecule inhibitor I-BRD9 can inhibit this process.

Article Snippet: I-BRD9 was purchased from MedChemExpress, and 740Y-P (PI3K activator, #HY-P0175; 20 μM, treatment time 24 h) was purchased from MedChemExpress, and all the reagents were dissolved in DMSO.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: BRD9 Inhibition, Alone or in Combination with Cytostatic Compounds as a Therapeutic Approach in Rhabdoid Tumors

doi: 10.3390/ijms18071537

Figure Lengend Snippet: Half maximal inhibitory concentrations (IC 50 ) of BI-9564 and I-BRD9 in RT cell lines incubated for 72 or 144 h. Cell proliferation was evaluated by MTT cytotoxicity assays. ( n ≥ 3).

Article Snippet: I-BRD9 (Tocris Bioscience, Bristol, UK) and BI-9564 (Tocris Bioscience, Bristol, UK) were dissolved in Dimethyl Sulfoxide (DMSO) (AppliChem, Hannover, Germany) as stock solutions of 10 mM and aliquoted to only freeze thaw once.

Techniques: Incubation

Journal: International Journal of Molecular Sciences

Article Title: BRD9 Inhibition, Alone or in Combination with Cytostatic Compounds as a Therapeutic Approach in Rhabdoid Tumors

doi: 10.3390/ijms18071537

Figure Lengend Snippet: Percentage of cells in G1 cell cycle phase after incubating with BI-9564 and I-BRD9 at the indicated concentrations for 72 h. ( n ≥ 3; mean ± SD; * treatment vs. control with p < 0.05, one-way ANOVA).

Article Snippet: I-BRD9 (Tocris Bioscience, Bristol, UK) and BI-9564 (Tocris Bioscience, Bristol, UK) were dissolved in Dimethyl Sulfoxide (DMSO) (AppliChem, Hannover, Germany) as stock solutions of 10 mM and aliquoted to only freeze thaw once.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: BRD9 Inhibition, Alone or in Combination with Cytostatic Compounds as a Therapeutic Approach in Rhabdoid Tumors

doi: 10.3390/ijms18071537

Figure Lengend Snippet: Percentage of dead RT cells after incubation with different BI-9564 and I-BRD9 concentrations for 72 h. ( n ≥ 3; mean ± SD, * treatment vs. control with p < 0.05, one-way ANOVA).

Article Snippet: I-BRD9 (Tocris Bioscience, Bristol, UK) and BI-9564 (Tocris Bioscience, Bristol, UK) were dissolved in Dimethyl Sulfoxide (DMSO) (AppliChem, Hannover, Germany) as stock solutions of 10 mM and aliquoted to only freeze thaw once.

Techniques: Incubation, Control

Journal: International Journal of Molecular Sciences

Article Title: BRD9 Inhibition, Alone or in Combination with Cytostatic Compounds as a Therapeutic Approach in Rhabdoid Tumors

doi: 10.3390/ijms18071537

Figure Lengend Snippet: IC 50 and combination indices (CI) of combined treatments. Treatment of BT12 and G401 with combinations of I-BRD9 and three cytotoxic drugs for 72 h. CI < 1 indicates synergistic effects and CI > 1 antagonistic effects of combined drugs. R 2 represents the determination coefficient of linear regression in median effect plot. ( n ≥ 3).

Article Snippet: I-BRD9 (Tocris Bioscience, Bristol, UK) and BI-9564 (Tocris Bioscience, Bristol, UK) were dissolved in Dimethyl Sulfoxide (DMSO) (AppliChem, Hannover, Germany) as stock solutions of 10 mM and aliquoted to only freeze thaw once.

Techniques: