Ara c

The cells used in this study were authenticated by STR profiling and have been tested for mycoplasma contamination. ARPE-19 or 661W cells were cultured in DMEM/F12 containing 10% fetal bovine serum and 1% penicillin-streptomycin. For Ara-C treatment, 661W cells were seeded on cover slides and treated with 10 μΜ of Ara-C (MedChemExpress, #HY-13605) for 12 h, and then the cells were either treated with or without JQ1 (1 μΜ) for 6 h or left untreated. cells were then fixed with 4% paraformaldehyde and subjected to IF analysis. To induce cytosolic DNA leakage, 661W cells were treated with or without 600 μM H₂O₂ for 2 h and then allowed to recover for 3 days before IF analysis. To determine the effect of BRD4 inhibitors on cGAS-STING transcription, 661W Cells were pretreated with 10 μM of JQ1, I-BET or OTX for 24 h, then subjected to IR (8 Gy) and harvested 24 h post-treatment. Alternatively, 661W cells were treated with Ara-C (10 μM) for 24 h and then BRD4 inhibitors (10 μM) were added for additional 48 h before analysis. To determine the autophagy of JQ on cytosolic DNA, 661W Cells were treated with DMSO, JQ1 (1 μM, 18 h) or Ara-C (10 μM, 18 h) as indicated. For Ara-C + JQ1 group, cells were pretreated with 10 μM of Ara-C, and then 1 μM of JQ1 was added in the presence of Ara-C for another 6 h.

HL60, THP-1, MV4-11, HEL, K562, and 293 T cells were obtained from Shanghai Institute of Cell Science, Chinese Academy of Sciences. All cell lines were cultured in RPMI 1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS and were maintained at 37 °C in 5% CO₂. All cells were subjected to short tandem repeat profiling and an incubation period of no more than two months. Ara-C, adriamycin (ADM), DDP, paclitaxel, 5-fluorouracil (5-FU), and methotrexate (MTX) were purchased from MedChemexpress CO., Ltd. Ara-C-resistant HEL (HLE-R) and HL60 (HL60-R) cells were generated by culturing cells in medium with a gradient of progressively increasing drug concentrations, with a final concentration of 8 μM. For all assays, drug-resistant cells were cultured for 48 h in drug-free medium before use in experiments.

AraC and CR-1-31-B were obtained from MedChemExpress and resuspended in DMSO. Venetoclax was obtained from Selleckchem and resuspended in DMSO. Aliquots of stock solution of the drugs were stored at − 80 °C and diluted into working solutions immediately before use. MOLM-14 cells were plated at 0.5× 10⁶ cells/ml and treated with drug at the indicated concentrations or an equal volume of vehicle.