Chir 124 s2683

Tumor tissue samples with their matched normal tissue were obtained from the International DIPG/DMG Registry as described previously [34 (link)]. Briefly, patients with DIPG were informed and consented to the IRB-approved protocol (Study ID: 2016-3357) at Cincinnati Children’s Hospital Medical Center. Diagnosis of DIPG was based on clinical symptoms and imaging characteristics on pretreatment magnetic resonance imaging (MRI). Primary normal human foreskin fibroblasts (HFF) strains (ATCC CRL-2091), human cervical carcinoma cell lines (HeLa), and the human osteosarcoma cell lines (Saos-2) (ATCC HTB-85) were purchased from ATCC. HFF and HeLa cells were cultured in DMEM (Gibco) supplemented with 10% FBS, while Saos-2 cells were cultured in McCoy’s 5A medium supplemented with 15% FBS. Pediatric GBM cell lines, CCHMC-DIPG-1, R0315-GBM, SJ-HGGX42 and HSJD-GBM002, were cultured in neurosphere stem cell media as described elsewhere [35 (link),36 (link)]. KNS42 cells were cultured in DMEM (Gibco) supplemented with 10% FBS. KNS42 cells were obtained from the JCRB (Japan Cancer Research Resources) cell bank. HSJD-GBM002 cells were generous gifts from Dr. Angel Montero Carcaboso. SJ-HGGX42 cells were obtained from PBTP (https://pbtp.stjude.cloud). ATR inhibitor VE-822 (S7102) and CHK1 inhibitor (Chir-124) (S2683) were purchased from Selleckchem.