Extended Data Fig. 6: UPP1, UPP2 and MITF expression across the CCLE collection and in melanoma.
From: Salvage of ribose from uridine or RNA supports glycolysis in nutrient-limited conditions
(a) UPP1, UPP2 and MITF expression across the complete CCLE collection (n = 30 lineages). (b) Protein immunoblot of a panel of melanoma (n = 9) and non-melanoma (n = 3, 293T, K562 and HeLa) cell lines grown and showing expression of UPP1 as well as MITF, TYR and MLANA, three melanoma markers. MDA: MDA-MB-435S. (c) Top: Immunoblot of UACC-257 melanoma cells with wild-type (UPP1WT) and knock-out (UPP1KO). Bottom: Immunoblot of MDA-MB-435S melanoma cells in wild-type (UPP1WT), knock-out (UPP1KO) and hypomorphic (UPP1hypo) clones (see methods). (d) Cell growth assay of MDA-MB-435S clones in sugar-free media containing dialyzed FBS complemented with 10 mM of either glucose or uridine and dialyzed FBS. Negative doublings indicate cell death. Data are shown as mean ± SEM with two-sided t-test relative to UPP1WT cells in the same media (n = 3 replicate wells, P < 2.9 × 10−6, P < 1.3 × 10−5, P < 3.1 × 10−7). (e) Cell growth assay of three melanoma cell lines with high UPP1 expression in sugar-free media complemented with 10 mM of glucose or 0.5 mg/mL of RNA. Data are shown as mean ± SEM with two-sided t-test relative to UPP1WT and were not corrected for multiple comparison (n = 3 replicate wells, P < 4.5 × 10−3, P < 1.3 × 10−4, P < 1.5 × 10−3. TUBB and Actin loading controls were performed on the same gels. All growth assays and screens included 4mM L-glutamine and 10% dialyzed FBS.
