Zotatifin

Caki-2 cells were cultured in McCoy’s 5A (modified) medium supplemented with 10% FCS, Calu-3 cells were cultured in MEM with 10% FCS and 1x non-essential amino acids and both obtained from ATCC (Manassas, VA, USA). HEK293T cells were cultured in DMEM supplemented with GlutaMAX, 10% heat-inactivated FCS. Caco-2 cells were from Sigma Aldrich (Schnelldorf, Germany) and were cultured in EMEM medium supplemented with 10% FCS, L-glutamine and non-essential amino acids (M7145, Sigma Aldrich, Schnelldorf, Germany). HepG2 was obtained from Sigma Aldrich (Schnelldorf, Germany) and cultured in DMEM supplemented with GlutaMAX and 10% heat-inactivated FCS. All media contained 1% penicillin/streptomycin and the cells were cultured at 37 °C in a 5% CO₂ atmosphere. Silvestrol was dissolved in DMSO and further diluted in media (c_stock = 6 mM, maximal DMSO concentration during experiments 0.1% v/v). EGTA, famotidine and carbamazepine were from Sigma Aldrich (Schnellendorf, Germany). Lucifer Yellow (sc-215269) was obtained from Santa Cruz. Celecoxib was synthesized by WITEGA Laboratorien Berlin-Adlershof GmbH (Berlin, Germany). Forskolin and ionomycin were obtained from Sigma Aldrich (Schnelldorf, Germany). Silvestrol was provided by the Sarawak Biodiversity Centre (Kuching, North-Borneo, Malaysia; purity > 99%) and zotatifin was bought from MedChemExpress (USA, purity: 98%).

Silvestrol was obtained from the Sarawak Biodiversity Centre (Kuching; North-Borneo, Malaysia; purity > 99%). A 6 mM stock solution was prepared in DMSO (sterile-filtered; Roth). Zotatifin (MedChemExpress, Monmounth Junction, NJ, USA; purity: 98%), CR-1-31-B (also known as CR-31-B (−)) and the inactive enantiomer CR-1-30-B (also known as CR-31-B (+), for further details see [20 (link),31 (link),32 (link),33 (link)]) were dissolved in DMSO at a concentration of 10 mM. All stock solutions were stored at −20 °C and diluted in the corresponding growth media (DMEM or IMDM).