Green fluorescent protein and firefly luciferase-positive AsPC-1 cells (AsPC-1 WT) were generated in-house as described previously by Schaefer et al.28 (link) Luciferase expressing Raji cells were generated as described previously by Seitz et al.27 (link) AsPC-1 WT secreting latent TGF-β (AsPC-1-TGF) were generated via transduction of AsPC-1 WT with lentiviral vectors (LVV) encoding human TGF-β (UniProtKB-A0A024R0P8-1). The plasmid encoding human TGF-β was kindly provided by Stefan Edelburg (Miltenyi Biotec, Bergisch-Gladbach, Germany). Secretion of TGF-β was confirmed by ELISA (BioLegend, catalog no.: 432907). Human embryonic kidney-293 T cells (HEK293T, DSMZ no: Acc635) were obtained from DSMZ.
AsPC-1 WT and AsPC-1-TGF were cultured in RPMI 1640 (Biowest catalog no.: L0501) supplemented with 2 mM glutamine (Lonza, catalog no.: BE17-605E) and 10% fetal bovine serum (EXIMUS Maximus FBS, Catus Biotech, catalog no.: BS-2020-500). HEK293T cells were cultured in Dulbecco’s modified Eagle medium (Biowest, catalog no.: L0104) supplemented with 10% FBS. Cell lines and primary human cells were cultured at 37°C and 5% CO2.
AsPC-1 WT and AsPC-1-TGF were cultured in RPMI 1640 (Biowest catalog no.: L0501) supplemented with 2 mM glutamine (Lonza, catalog no.: BE17-605E) and 10% fetal bovine serum (EXIMUS Maximus FBS, Catus Biotech, catalog no.: BS-2020-500). HEK293T cells were cultured in Dulbecco’s modified Eagle medium (Biowest, catalog no.: L0104) supplemented with 10% FBS. Cell lines and primary human cells were cultured at 37°C and 5% CO2.
