Cfpac 1

PDAC cell lines (AsPC-1, BxPC-3, CFPAC-1, MIAPaCa-2 and PANC-1), human pancreatic ductal immortalized cells (HPNE) and human embryonic kidney cells (HEK-293T) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). BxPC-3 and CFPAC-1 cells were cultured in RPMI 1640 medium (Gibco). AsPC-1, PANC-1, MIA PaCa-2, HPNE and HEK-293T cells were cultured in high-glucose DMEM (Gibco). All media were supplemented with 10% fetal bovine serum (FBS, Gibco), 100 IU/mL penicillin and 100 μg/mL streptomycin (NCM, Suzhou, China). All cells were cultured at 37 °C in a humidified atmosphere containing 5% CO₂. All cells were authenticated using STR profiling and tested negative for mycoplasma contamination.

The human pancreatic cancer cell lines PANC-1 and CFPAC-1 used in our study were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and stably cultured, passaged, and cryopreserved in our laboratory. Basic Dulbecco's modified Eagle's medium (DMEM, Gibco, USA) with 10% fetal bovine serum (FBS, Sigma, St. Louis, MO, USA) was utilized to culture PANC-1 cells, while basic RPMI-1640 medium (Gibco, USA) with 10% FBS (Sigma, St. Louis, MO, USA) was utilized to culture CFPAC-1 cells, which were cultured in an incubator at 37°C, 5% CO₂, and 95% humidity. The drug curcumin was purchased from MedChemExpress (MCE, New Jersey, USA). The anti-sestrin2 antibody was purchased from Proteintech (Chicago, USA), while the anti-Nrf2 antibody, p-Nrf2 antibody, Keap1 antibody, HO-1 antibody, and NQO-1 antibody were purchased from Abcam (Britain).

The normal immortalized human pancreatic epithelial cell line (HPDE6C7) and four human pancreatic cancer cell lines (CFPAC‐1, BXPC3, L3.6pl and Panc‐1) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). All the relevant cells were cultured in DMEM (Sigma‐Aldrich, St. Louis) supplemented with 10% fetal bovine serum (FBS; HyClone, Logan, USA) and kept in a humidified incubator with 5% carbon dioxide at 37°C. The pcDNA3.1‐SNHG14 and pcDNA3.1‐ANXA2 (vector with SNHG4 or ANXA2 overexpression) and empty vectors were from GenePharma (Shanghai, China). The miR‐613 mimic and its negative control (NC), and SNHG14 small interfering RNA (siRNA) (si‐SNHG14) and scrambled siRNA for SNHG14 (si‐NC) were from RiboBio (Guangzhou, China). For cell transfections, cells were grown on 6‐well plates until 60% confluence, and then cells were transfected by miRNA, siRNA, or plasmid using Lipofectamine 2000 reagent (Invitrogen, Carlsbad).

BxPC-3, CFPAC-1, and PANC-1 human PDAC cell lines were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China), and the immortal human pancreatic duct epithelial cell line HPDE6-C7 was provided as a gift from Professor Dongxin Lin from SYSUCC. Cells were cultured at 37°C in a humidified atmosphere of 5% CO₂. The culture medium was used as recommended and was supplemented with 10% fetal bovine serum (Gibco, California, USA).
Cell lines with SYPL1 stably silenced or overexpressed were established using lentiviruses (iGeneBio, GuangZhou, China). We also used siRNA (RiboBio, Guangzhou, China), which was transfected into cells by Lipofectamine 2000 (Invitrogen, California, USA) according to the manufacturer's instructions. To knockdown SYPL1, the following validated target sequence was used: CCTCATAGGCGATTACTCT.

The human pancreatic cancer cell lines (BXPC3, CFPAC-1, Panc-1 and L3.6pl) and HEK-293T cells were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Sigma, St Louis, USA) supplemented with 10% fetal bovine serum (HyClone, GE Healthcare Life Science, Logan, USA) and incubated in a humidified chamber supplemented with 5% CO₂ at 37°C.

Human pancreatic cancer AsPC-1, BxPC-3, CAPAN-1, CFPAC-1, PANC-1, and SW1990 cell lines were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). AsPC-1 and BxPC-3 cells were cultured in RPMI 1640 culture medium with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S), while the others were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS and 1% P/S. The culture conditions were 37 °C and a 5% CO₂ atmosphere. For studies in hypoxia, cells were grown at 37 °C in an atmosphere of 1% O₂. For the cellular function assay, all PDAC cells were cultured in medium with 5 mM glucose and 2 mM L-glutamine in the absence of FBS. Experiments in Figs. 4c–f, 5c and 7b–f were performed with cells cultured in hypoxia condition to better simulate the hypovascular tumor microenvironment in PDAC. Cell function assays performed in non-hypoxia conditions were shown in Figs. S1–S3.