Journal: Cell Death & Disease

Article Title: Therapeutic targeting of ARID1A-deficient cancer cells with RITA (Reactivating p53 and inducing tumor apoptosis)

doi: 10.1038/s41419-024-06751-1

Figure Lengend Snippet: A Interaction between ARID1A and p53. Coimmunoprecipitation assays were performed in HCT116 cell lines as described in “Methods” section. Input (cell lysates without immunoprecipitation) and IgG served as positive and negative controls, respectively. B RT-qPCR analysis of CDKN1A , PUMA , and NOXA mRNA level in HCT116 ARID1A +/+ and ARID1A − / − clones. ** P < 0.01, one-sample t -test. C Upregulation of p53, PUMA, and NOXA levels and downregulation of p21 level in HCT116 ARID1A − / − cell lines. D Downregulation of p53, PUMA, and NOXA levels and upregulation of p21 level in RKO ARID1A OE cell lines. E Upregulation of p53, PUMA, and NOXA levels and downregulation of p21 level by ARID1A silencing. F Downregulation of p53, PUMA, and NOXA levels and upregulation of p21 level by ARID1A overexpression. G Upregulation of p53, p21, PUMA, and NOXA levels with the increasing concentration of RITA treatment for 48 h in HCT116 cells. H Upregulation of p53, p21, PUMA, and NOXA level by 100 nM RITA treatment for 48 h in HCT116 ARID1A +/+ and ARID1A − / − cell lines. I , J Effect of RITA on the cell cycle progression of HCT116 ARID1A − / − cells. Cells were treated with 100 nM RITA for 48 h before flow cytometry. Cell cycle distribution ( I ) and cell population quantification ( J ) are shown. Error bars represent s.d. ( n = 3 independent experiments). ANOVA ** P < 0.01. K , L Effect of RITA on the cell cycle progression of RKO ARID1A − / − cells. Cells were treated with 10 μM RITA for 48 h before flow cytometry. Cell cycle distribution ( K ) and cell population quantification ( L ) are shown. Error bars represent s.d. ( n = 3 independent experiments). ANOVA ** P < 0.01.

Article Snippet: Each aliquot of protein sample was run on an SDS-polyacrylamide gel electrophoresis and transferred onto a nitrocellulose membrane for immunoblotting with primary antibodies, including ARID1A (Cell Signaling Technology, #12354S, 1:2000 dilution), Cleaved PARP (Cell Signaling Technology, #5625S, 1:2000 dilution), Cleaved caspase3 (Cell Signaling Technology, #9661S, 1:500 dilution), MDM2 (Cell Signaling Technology, #86934S, 1:1000 dilution), p53 (Cell Signaling Technology, #2527S, 1:2000 dilution), p21 (Cell Signaling Technology, #2947S, 1:2000 dilution), PUMA (Cell Signaling Technology, #98672S, 1:2000 dilution), NOXA (Cell Signaling Technology, #14766S, 1:2000 dilution), Phospho-ATM (Ser1981) (Cell Signaling Technology, #5883T, 1:1000 dilution), Phospho-ATR (Ser428) (Cell Signaling Technology, #2853T, 1:1000 dilution), Phospho-BRCA1 (Ser1524) (Cell Signaling Technology, #9009T, 1:1000 dilution), Phospho-Chk1 (Ser345) (Cell Signaling Technology, #2348T, 1:1000 dilution), Phospho-Chk2 (Thr68) (Cell Signaling Technology, #2197T, 1:1000 dilution), Phospho-p53 (Ser15) (Cell Signaling Technology, #9286T, 1:1000 dilution), Phospho-Histone H2A.X (Ser139) (Cell Signaling Technology, #9718T, 1:1000 dilution), MDM4 (Proteintech, #28747-1-AP, 1:1000 dilution), FANCD2 (Proteintech, # 28619-1-AP, 1:1000 dilution) and β-actin (Cell Signaling Technology, #3700S, 1:4000 dilution) antibodies, followed by horseradish peroxidase-conjugated secondary antibodies (ZEN-Bioscience, 511203).

Techniques: Immunoprecipitation, Quantitative RT-PCR, Clone Assay, Over Expression, Concentration Assay, Flow Cytometry

Journal: Cell reports

Article Title: In vivo RNA-seq and ChIP-seq analyses show an obligatory role for the C terminus of p53 in conferring tissue-specific radiation sensitivity

doi: 10.1016/j.celrep.2023.112216

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit monoclonal antibody to PUMA , Cell Signaling , 14570S; RRID:AB_2798517.

Techniques: Recombinant, Protein Extraction, Membrane, Staining, Protease Inhibitor, Plasmid Preparation, TUNEL Assay, Purification, ChIP-qPCR, Software