Jurkat

Human cancer cell lines HL-60, K562, Jurkat, HeLa, A549, MCF-7, HepG2, A431, and MIA PaCa-2, along with human normal cell lines WI-38 and 1C3D3, were obtained from the RIKEN Cell Bank (RIKEN BRC, Tsukuba, Japan). Human normal cell line YS-1 was obtained from the JCRB Cell Bank (Osaka, Japan). Human cancer cell lines U937, PC-3, DLD-1, Hep3B, WM266-4, SK-MEL-28, HT-1080, and BxPC-3 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). HL-60, K562, Jurkat, U937, MCF-7, DLD-1, and BxPC-3 cells were cultured in RPMI 1640 (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal calf serum (Sigma-Aldrich), 50 units/mL penicillin G (Gibco), and 50 μg/mL streptomycin (Gibco). HeLa, A549, PC-3, HepG2, Hep3B, WM266-4, SK-MEL-28, HT-1080, A431, MIA PaCa-2, and WI-38 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) containing 10% fetal calf serum, 50 units/mL penicillin G, and 50 μg/mL streptomycin. YS-1 cells were cultured in DMEM containing 5% fetal calf serum, 50 units/mL penicillin G, and 50 μg/mL streptomycin. 1C3D3 cells were cultured in DMEM containing 5% fetal calf serum, 10% newborn bovine serum (SAFC Biosciences, Lenexa, KS, USA), 2.5% horse serum (Gibco), 50 units/mL penicillin G, and 50 μg/mL streptomycin. All cell lines were incubated at 37°C in a humidified atmosphere containing 5% CO₂.

The human cell lines JURKAT (Clone E6-1) and MOLT-3 were purchased from ATCC. The human cell lines DND-41, HPB-ALL, ALL-SIL, and PEER were purchased from DSMZ. JURKAT, DND-41, and ALL-SIL were cultured in 90% RPMI-1640 Glutamax (Gibco), supplemented with 10% of heat-inactivated fetal bovine serum (FBS, Gibco) and 10,000 U/mL penicillin and streptomycin (Invitrogen). HPB-ALL and PEER were cultured in 80% RPMI-1640 Glutamax, supplemented with 20% heat-inactivated FBS and 10,000 U/mL penicillin and streptomycin. All cell lines were cultured at 37 °C, in a humidified incubator with 5% CO₂, and routinely checked for mycoplasma, using the EZ PCR Mycoplasma detection Kit (Biological Industries); only mycoplasma-free cells were used. To avoid artefacts derived from long-term culture of immortalized cell lines, typically cells were not kept in culture for more than 2 months, starting from frozen stocks derived from the commercial repository.
Persister cells were generated by treating the DND-41 cells for up to 12 weeks with 0.1–1 μM GSI (GSI XXI, Compound E; Calbiochem). The GSI was chosen based on the original publication where DND-41 persister cells were generated for the first time^{14 (link)}.
Cell count was performed by trypan blue exclusion on the automated CytoSMART cell counter (Corning).

Cell lines including lung cancer cell lines HCC827 (CRL-2868), H1975 (CRL-5908), A549 (CCL-185), epidermoid carcinoma cell line A431 (CRL-1555), cervical carcinoma cell line Hela (CCL-2), breast cancer cell line MCF-7 (HTB-22) were purchased from the American Type Culture Collection (ATCC, Manassas, VA), lung cancer cell line PC-9 (RCB 4455) and acute T cell leukemia cell line Jurkat (RCB 3052) were obtained from RIKEN Cell Bank (Ibaraki, Japan). Cells were cultured in Dulbecco’s modified Eagle medium containing 10% fetal bovine serum (Gibco), penicillin (100 U/mL), and streptomycin (100 µg/mL). Jurkat cells were cultured in RPIM 1640 (Gibco). All cells were maintained in a humidified 37 °C incubator with 5% CO₂. All cell lines used in our study have been authenticated and the mycoplasma tests of the cell lines were negative. Details of the cellular experiments are provided in Supplementary Methods.

HEK293T, Jurkat, Hela, and PANC-1 cell lines were acquired from the Iranian Biological Resource Center (IBRC). HEK293T, Hela, and PANC-1 cells were maintained in D10 media, comprising DMEM (Gibco, Life Technologies), 10% fetal bovine serum (FBS), and 1% penicillin/streptomycin (Gibco, Life Technologies). Jurkat cells were cultured in R10 media, containing RPMI-1640 (Gibco, Life Technologies) supplemented with 10% FBS, 25 mM HEPES (Sigma Aldrich), 2 mM glutamine (Gibco), and 1% penicillin/streptomycin. Flow cytometry was used to validate mesothelin expression in the relevant cell lines prior to experiments. Regular mycoplasma contamination checks were conducted on all cell lines.

THP1, Jurkat, Hela, L929 and RAW264.7 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). THP1-Lucia ISG cells were purchased from Invivogen. cGAS^-/- Hela cells were a kind gift from Dr. Zhengfan Jiang (School of Life Science, Peking University, Beijing, China). Human peripheral blood mononuclear cells (PBMCs) were purchased from LDEBIO (Guangzhou, China). THP1, THP1-Lucia ISG cells, Jurkat and PBMCs were cultured in RPMI 1640 (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gbico) and 0.1% Normocin (InvivoGen, Cat# ant-nr) and 0.1% mycoplasma elimination reagent (Yeasen, Cat# 40607ES03) at 37 °C with 5% CO₂. Hela, cGAS^-/- Hela, L929 and RAW264.7 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gbico) supplemented with 10% FBS and 0.1% Normocin (InvivoGen, Cat# ant-nr) and 0.1% mycoplasma elimination reagent (Yeasen, Cat# 40607ES03) at 37 °C with 5% CO₂.