Capan 2

Normal pancreatic duct epithelial cells (HPNE) and human PC cell lines (PANC-1, SW1990, AsPC-1, Capan-2) were acquired from the Shanghai Cell Research Institute (Shanghai, China). HPNE and AsPC-1 cells were cultured in 1640 medium (Gibco, USA), and PANC-1, SW1990, and Capan-2 cells were cultured in DMEM medium (Gibco), both of which contained 10% fetal bovine serum (FBS; Gibco) and 1% of penicillin-streptomycin (Gibco), and were incubated in an incubator with 37°C and 5% CO₂.

The pancreatic cancer cell lines (Pan02, MIA PaCa-2, BxPC-3, PANC-1, Panc-28, SW1990, Capan-2) were obtained from ATCC (Manassas, VA, USA). CHO and HEK293 were purchased from the National Infrastructure of Cell Line Resource (Shanghai, China). HEK293, Pan02, BxPC-3, PANC-1 and Panc-28 were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) with 100 U/mL of penicillin and 100 µg/mL of streptomycin (Gibco, Carlsbad, CA, USA). MIA PaCa-2 was grown in DMEM supplemented with 10% FBS and 2.5% horse serum (Gibco, Carlsbad, CA, USA). SW1990 was maintained in L15 medium supplemented with 10% FBS. Capan-2 was cultured in RPMI-1640 (Gibco, Carlsbad, CA, USA) supplemented with 10% FBS. CHO was grown in DMEM-F12 (Gibco, Carlsbad, CA, USA) supplemented with 10% FBS. All cells were maintained at 37 °C in a humidified 5% CO₂ atmosphere. Cell lines were identified by short tandem repeat analysis and checked for mycoplasma contamination.

The human pancreatic cancer cell lines PANC-1, MIA PaCa-2, and SW1990 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA); AsPC-1, BxPC-3, Capan-2, and Panc 03.27 cells were purchased from the Cell Repository of the Chinese Academy of Sciences (Shanghai, China). The immortalized HPDE cell line was obtained from the Beijing North Carolina Chuanglian Biotechnology Research Institute (Beijing, China). Capan-2, MIA PaCa-2, and PANC − 1 cells were grown in Dulbecco Modified Eagle Medium (Gibco, Carlsbad, CA, USA) supplemented with 10% FBS (Gibco), 100 U/mL penicillin G, and 100 mg/mL streptomycin (Sigma-Aldrich, St. Louis, MO, USA). AsPC-1, BxPC-3, Panc 03.27, and HPDE were grown in 1640 medium (Gibco) supplemented with 10% FBS (Gibco), 100 U/mL penicillin G, and 100 mg/mL streptomycin (Sigma-Aldrich). All cells were grown at 37 °C in a humidified 5% CO₂ incubator. The reagents and antibodies used in this study are listed in the Additional file 10: Table S5. Reagents and antibodies.

Pancreatic cancer cell lines Panc1, MiaPaCa2, BxPC3, Capan‐2 (Human origin), and Panc02 (Mouse C57BL/6 origin) were obtained from American Type Culture Collection (ATCC; Manassas, VA, USA) and Frederick National Laboratory for Cancer Research (Frederick, MD, USA), respectively. The Panc1, MiaPaCa2, and Panc02 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, 4.5 g/L glucose; Gibco, Rockville, MD, USA) supplemented with 10% heat‐inactivated fetal bovine serum (FBS; Gibco), 1 mM sodium pyruvate (Cat. P5280, Sigma‐Aldrich, St. Louise, MO, USA), 100 U/mL penicillin (Gibco) and 100 mg/mL streptomycin (Gibco). BxPC3 and Capan‐2 were cultured in RPMI‐1640 medium (Gibco) and McCoy's 5A Medium (Gibco), respectively, supplemented with 10% FBS (Gibco). All cells were maintained in a humidified incubator containing 5% CO₂ at 37°C.
Berberine chloride (C20H18ClNO4, PubChem CID: 12456) was purchased from Sigma‐Aldrich (Cat. B3251‐10G, Lot: #SLBG1303). For AMPK inhibitor and mTOR inhibitor treatment, cells are treated with compound c (20 μΜ; PubChem CID: 11524144, Sigma‐Aldrich), Rapamycin (10 nM; PubChem CID: 5284616, Cat. R0395, Sigma‐Aldrich), 7ACC2 (10 μΜ; PubChem CID: 72696735, Cat. HY‐D0713, MedChemExpress), respectively, for 24 hours. For lactate treatment, cells are treated with L‐lactate (10 mM; PubChem CID: 107689, Cat. L1750, Sigma‐Aldrich) for 48 hours.

Pancreatic cell line hTERT-HPNE and pancreatic cancer cell lines (PANC-1, SW1990, BxPC-3, MIA PaCa-2, Capan-2, and HPAF-II) were purchased from the Cell Bank of The Chinese Academy of Sciences and incubated in medium containing 10% fetal bovine serum (FBS, Excell Bio, CN) at 37 °C under 5% CO₂. The validation and authentication for the cell lines had been performed. SW1990, BxPC-3, and Capan-2 cells were plated in RPMI-1640 (Gibco, Waltham, MA, USA). PANC-1, MIA PaCa-2, HPAF-II, and hTERT-HPNE cells were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Waltham, MA, USA).
The human siRNAs targeting PPM1K (siRNA-1, siRNA-2, and siRNA-3) and negative control (NC) were purchased from Kidan Biosciences (Guangzhou, China). For stable transfection, cells were plated in six-well plates 24 h in advance, then transfected with siRNAs using lipofectamine 2000 reagents (Invitrogen, Waltham, MA, USA) according to the manufacturer’s instructions. Two days after transfection, these cells were harvested for the following experiments. The siRNA sequences are listed in Supplementary Table S1.

The pancreatic epithelial cell lines HPDE and PC (AsPc-1, BXPC-3, Capan-2, CFPAC-1, PANC-1, MIA PaCa-2, and SW1990) were obtained from the American Type Culture Collection. HPDE, AsPc-1, and BXPC-3 cells were maintained in RPMI-1640 medium (Gibco) containing 10% fetal bovine serum (FBS; Gibco), while Capan-2, CFPAC-1, PANC-1, MIA PaCa-2, and SW1990 were cultured in DMEM containing 10% FBS. Both cell lines were cultured at 37°C in a humidified 5% CO2 incubator according to ATCC protocols.

K562 (ATCC) cells were cultured in IMDM (Gibco, Cat. No. 12440053) supplemented with 10% FCS (TICO Europe, Cat. No. FBSEU500). Capan-2 (ATCC) cells were cultured in RPMI-1640 (Gibco, Cat. No. 61870036) supplemented with 15% FCS (TICO Europe, Cat. No. FBSEU500). MiaPaca-2 (ATCC) cells were cultured in DMEM (Gibco, Cat. No. 31885023) supplemented with 15% FCS (TICO Europe, Cat. No. FBSEU500) and 2.5% horse serum (Gibco, Cat. No. 16050130). All cells were maintained at 37°C in a humidified incubator containing 21% O₂ and 5% CO₂ (Sanyo Electric).