Ar42j

All cell culture materials were purchased from Fisher Scientific unless otherwise noted. The SSTR2-positive rat pancreatic cancer cell line AR42J (ATCC, Manassas, VA, USA) was cultured in Gibco™ IMDM media supplemented with 10% fetal bovine serum (FBS) and 0.1% gentamicin. Cells were incubated at 37 °C in a 5% CO₂ atmosphere and were harvested using 0.05% trypsin-EDTA. Cells were washed once with supplemented IMDM prior to use. Cells were counted with a Nexcelom (Lawrence, MA, USA) auto T4 bright field cell counter.

AR4-2J was bought from ATCC (Rockville, MD, USA) and cultured in DMEM/F12 supplemented with 20% fetal bovine serum in a CO₂ incubator under humidified atmosphere (5% CO₂/95% air) at 37 °C, as reported before [1 (link),2 (link),28 (link),33 (link),52 (link)].
Solid E. coli medium LB/Kana and LB/Amp were sterilized and culture plates made. Liquid E. coli medium LB/Kana and LB/Amp had the same composition, but without agar.

AR42J (from ATCC) were cultured in F-12k medium with 20% heat-inactivated FBS and 1% penicillin and streptomycin. PC3 (from ATCC), PC3-PIP (from NIBIB at NIH) and 4T1-fluc cell lines (from ATCC) were cultured in RPMI-1640 medium with 10% heat-inactivated FBS and 1% penicillin and streptomycin. Cells were grown in a humidified atmosphere (5% CO₂, 37 °C). All cells were tested to be free of mycoplasma.

AR4-2J was bought from ATCC (Rockville, MD, United States) and cultured in DMEM/F12 supplemented with 20% fetal bovine serum (Hyclone) and antibiotics in a CO₂ incubator under 5% CO₂ at 37°C as before (Matthews and Cui, 1990a (link),b (link); Cui, 1998 (link); Duan et al., 2011 (link); Liang et al., 2013 (link)). HEK293 and CHO-K1 were purchased from Shanghai Institutes of Life Sciences Chinese Academy of Sciences and cultured in DMEM/F12 similarly.
Solid E. coli medium LB/Kana was sterilized and culture plates made. Liquid E. coli medium LB/Kana had the same composition but without agar.

Rat pancreatic acinar cell, AR42J (#CRL-1492, ATCC), was used in this study. Cell adherence culture at 5 cm × 5 cm cell culture bottle. RPMI 1640 medium was used for cell culture, in which 10% FBS and Antibic Antimycotic double antibody were added. The culture condition was 37 °C and the CO₂ concentration was 5%.
1 × 10⁶ cells were cultured in a 6-well plate for 24 h. Cerulein was added to medium with the final concentration of 10 nmol/L. After 4 h, TFC (50 mg/L) was added to culture cells for 24 h. Then, the supernatant of cell culture and AR42J cells were collected for subsequent experiments.

Pancreatic acinar cells AR4-2J, purchased from ATCC (USA), were cultured in Dulbecco’s Modified Eagle Medium/F12 supplemented with 20% fetal calf serum and antibiotics, and cultured in a 37 °C, 5% CO₂ incubator (American ThermoFisher Scientific Corporation). Cells in the logarithmic phase were trypsinized, and inoculated in 6-well plates at 1 × 10⁵ cells per well. After 24 h of routine culture, cell transfection was performed according to Lipofectamine 2000 (Invitrogen) when cell confluence reached 75%. The cultured cells were transfected with the following plasmids: sh-ATF4, sh-HDAC1, sh-NEP, oe-HDAC1, oe-KFL4 and corresponding controls. Silencing plasmids (pGPU6/Neo, C02003) were purchased from Shanghai GenePharma Co., Ltd, and overexpression plasmids (pCDNA3.1-FLAG-LPA2, P1224) were purchased from Wuhan Miaoling Biotechnology Co., Ltd. (Wuhan, China) Successfully transfected cells were incubated with 500 μM Na-TC for 12 h, and then treated with L-alanyl-L-glutamine (2 mM) and cerulein to construct a cellular AP model.