Cells, Cultivation, and Reagents

Human epithelial adenocarcinoma non-small cell lung cancer line A549 (ATCC^® CCL-185^TM) was used. Construction of the stable p21 knockdown A549 cells (A549^p21–) was described previously (Bunik et al., 2020b). The knock-down and mock control cells were maintained with puromycin at concentration of 0.2 μg/ml. The cells were cultivated in Dulbecco’s Modified Eagle’s medium (DMEM) (Thermo Fischer Scientific, 21855), which comprised 1 g/L glucose, 1 mM pyruvate, 4 mM GlutaMAX^TM, 10% FBS (Gibco, 10270), 100 U/ml penicillin and 0.1 mg/ml streptomycin (Thermo Fisher Scientific, 15140), at 37°C, 5% CO₂ in a humidified atmosphere.

The protease and phosphatase inhibitor cocktails cOmplete^TM (Roche, 04693116001) and PhosSTOP^TM (Roche, PHOSS-RO) were used. Cisplatin (PHR1624), NAD⁺ (N7004), NADP⁺ (N5755), isocitrate (I1252), nicotinamide (72340), ThDP (C8754), NADH (N8129), malic acid (240176), oxaloacetate (O4126), 2-oxoglutarate (K1750), CoA (C4282), sodium deoxycholate (D6750), IGEPAL CA-630 (I8896) were from Merck (Sigma-Aldrich).