Rpmi 1640 1x

Pan02 cells were provided by the Division of Cancer Treatment and Diagnosis (DCTD, NCI) and have been maintained in RPMI (1640-1X) (CORNING LOT NO. 16921011) supplemented with 10% fetal bovine serum, antibiotics (100 units/mL penicillin and 100 μg/mL streptomycin, Atlanta Biologicals). Pan02 cells, passage numbers between 4 and 6, were thawed for expansion, and cells with passage numbers between 10 and 20 were used in the in vitro experiments. Pan02 cells were passaged in T75 flasks with 0.5–1 million cells seeded into 16 ml of media and incubated it till 75% confluency was reached. Cell solution at a concentration of 5 × 10⁶ cells/mL was prepared for treatment.

Pan02 cells were provided by the Division of Cancer Treatment and Diagnosis (DCTD, NCI) and have been maintained in RPMI (1640-1X) (CORNING LOT NO. 16921011) supplemented with 10% fetal bovine serum, antibiotics (100 units/mL penicillin and 100 μg/mL streptomycin, Atlanta Biologicals). Pan02 cells, passage numbers between 4–6, were thawed for expansion, and cells with passage numbers between 10 and 20 were used in the in vitro experiments.

We cultured 5 Epstein-Barr Virus transformed lymphoblastoid cell lines (LCLs) at 37°C and 5% CO₂. These LCLs—GM18501, GM18504, GM19144, GM19239, and GM19153—were derived from the Yoruba (YRI) individuals from the International HapMap Consortium. The Coriell Cat #:Research Resource Identifiers (RRIDs) are GM18501:CVCL_P458, GM18504:CVCL_P460, GM19144:CVCL_P525, GM19239:CVCL_9634, and GM19153:CVCL_P531. These lines were authenticated and tested for mycoplasma contamination. Cell culture and RNA extraction were performed as described previously [52 (link)]. In brief, cells were grown in a glutamine depleted RPMI [RPMI 1640 1X from Corning (15–040 CM)] with 15% FBS, 2 mM GlutaMAX (from gibco (35050–061), 100 IU/mL Penicillin, and 100 ug/mL Streptomycin. The lines were passaged 3 times, maintained at 8×10⁵ cells, and grown to a concentration of 1×10⁶ cells per mL before RNA extraction, which was performed as described previously [52 (link)]. In brief, cells from each line were spun down and pelleted at 200 g at 500 RPM at 4°C for 2 min, washed with cold phosphate-buffered saline (PBS), and spun down again before aspirating the PBS. RNA was extracted using the miRNeasy kit (Qiagen) according to the manufacturer’s instructions, including the DNase step to remove potentially contaminating genomic DNA.