Hy 13982

Temozolomide (34219) was obtained from Sigma‐Aldrich. SC75741 (HY‐10496), SN50 (HY‐P0151), JSH‐23 (HY‐13982), and lomeguatrib (HY‐13668) were obtained from MedChemExpress. Primary antibodies against NF‐κB p65 (D14E12) (59674S, 1:1000), p‐NF‐κB p65 (Ser536) (93H1) (3033S, 1:1000), and IκBα (44D4) (4812, 1:1000) were purchased from Cell Signaling Technology. Primary antibodies against MGMT (ab69629) were purchased from Abcam. Primary antibodies against Lamin B1 (B‐10) (sc‐374015) were purchased from Santa Cruz Biotechnology. Primary antibodies against GAPDH (1A6) (MB001, 1:1000) were purchased from Bio‐World. Anti‐mouse IgG, HRP‐linked antibody, and anti‐rabbit IgG, HRP‐linked antibodies were purchased from Santa Cruz Biotechnology. Matrigel (354248) was obtained from BD Biosciences.

The chemicals used in vitro and in vivo assays are as follows: SB225002 (0.1 μM for the in vitro assays and 2 mg/kg for the mouse treatment, MedChemExpress; HY-16711), JSH-23 (5 μM for the in vitro assays and 2 mg/kg for the mouse treatment, MedChemExpress; HY-13982); RS504393 (0.2 μM for the in vitro assays, MedChemExpress; HY-15418); MG132 (Selleck Chemical; S2619s); cycloheximide (BioVision; 1041-1 G). SB225002 and JSH-23 in DMSO were diluted in corn oil for in vivo administration through intraperitoneal injection every other day or gavage daily, respectively. For immunotherapy, antibody intraperitoneal injection was started when subcutaneous tumor volume reached ~100 mm³ or Pten^PC−/−; Arid1a^PC−/− mice were 2.5-month-old. The following antibodies were injected alone or in combination: anti-mouse PD1 (BioXCell; Clone: RMP1-14, BE0146); anti-mouse CTLA4 (BioXCell; Clone: 9H10, BE0131); anti-mouse Ly6G (BioXCell; clone: 1A8, BE0075-1); and their respective isotype IgG controls. PD1/CTLA4 antibody were simultaneously administrated to the mice. Treatment was administered twice a week through intraperitoneal injections at a dosage of 200 μg/injection/antibody, and subcutaneous tumor volume was monitored every 4 days. CXCL2 was depleted from conditioned media by incubation with mouse antibody against Mip2/Cxcl2 (R&D Systems; MAB452).