Hs766t

Cell lines (PANC-1, Capan-1, HS766T, MIA-PaCa2, and CFPAC) were purchased from ATCC (Manassas, VA). HuR knockout clones were generated as previously described (17 (link),18 (link)). PDX line SPC_145 was a gift from Dr. Talia Golan (Sheba Medical Center) (19 (link)). All cell lines were tested monthly for mycoplasma and every 6 months STR validated. Cell lines for all experiments were below passage 20.

PDA cell lines (MIA PaCa-2, PANC-1, Capan-1, Hs 766T, PL11) were purchased from ATCC (Manassas, VA, 2012). All cell lines were routinely tested for mycoplasma using LookOut® Mycoplasma PCR Detection Kit (MP0035 SIGMA), and only early passage (<10) mycoplasma- negative cell lines were used for in vitro and in vivo experiments. As further validation, genomic DNA extracted, PCR amplified and sent for Sanger sequencing. All cell lines were validated as per the expected KRAS and p53 mutation status (21 (link)). Cells were cultured in standard DMEM media supplemented with 10% fetal bovine serum (FBS), 1% L-glutamine and 1% penicillin-streptomycin (Invitrogen) at 37°C and 5%CO₂. MIA PaCa-2 and Hs 766T with CRISPR/Cas9 knockout of HuR and MIA PaCa-2 cells with doxycycline inducible silencing of HuR were generated and characterized as previously described (18 (link), 22 ).

All the human cell lines (Hs766T, BxPc3, Patu8988T and Patu8902) used in this study were purchased from ATCC, used below passage 25 and continuously cultured in 100 U/ml penicillin and 100 U/ml streptomycin. The cell lines were authenticated by short tandem repeat (STR) profiling at the Institute for Applied Cancer Sciences, MD Anderson Cancer Center. The Patu8988T, Hs766T and Patu8902 cell lines were routinely cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS (Invitrogen). BxPc3 cell lines were routinely cultured in Roswell Park Memorial Institute (RPMI) 1640 (Invitrogen) with 10% FBS. Primary mouse cell lines were established in the laboratory (AK-B6, AK192, HY6468, PJAK4217, PJAK4298) as described previously^{34 (link)} and were routinely cultured in Roswell Park Memorial Institute (RPMI) 1640 (Invitrogen) 10% FBS (Invitrogen). For inducible Kras derived cell lines, 1 ug/ml of doxycycline was directly added to the media. For metabolic and metabolomic assays, 10% dialysed FBS (Atlanta Biologicals Inc.) was used. The cell lines were mycoplasma free, based on tests done monthly in the laboratory using Lonza’s MycoAlert Mycoplasma Detection Kit assays with confirmatory tests by PCR-based assays.