Extended Data Fig. 1: Characterization of YME1L-dependent spheroid growth. | Nature

Extended Data Fig. 1: Characterization of YME1L-dependent spheroid growth.

From: Lipid signalling drives proteolytic rewiring of mitochondria by YME1L

Extended Data Fig. 1

a, b, Spheroid surface area (a) and ATP levels (b) of HEK293 cell lines after 7 days of culture (a, n = 5 independent experiments; b, n = 3 independent experiments; # is number of spheroids per condition). c, d, Spheroid surface area (c) and ATP content (d) of WT and YME1L/ HeLa cells after 7 days (n = 3 independent experiments; # is number of spheroids per condition). e, Quantification of the NAD/NADH ratio in WT and YME1L/ HEK293 cells cultured in normoxia (21% O2) or hypoxia (0.5% O2) for 24 h (n = 3 independent experiments). f, Spheroid surface area of WT and YME1L/ HEK293 cells and of YME1L/ HEK293 cells expressing Lb-NOX (NADH oxidase from Lactobacillus brevis)47 after 7 days (n = 1 experiment; # is number of spheroids per condition). The expression of Lb-NOX was determined by SDS–PAGE and immunoblotting of control and Lb-NOX-expressing WT and YME1L/− HEK293 cells in normoxia (21% O2) or hypoxia (0.5% O2) for 24 h. g, The dependency of WT and YME1L/ HEK293 cells on glucose oxidation (left) and fatty acid oxidation (right) was monitored using the XF Fuel Flex Assay. Dependencies are calculated as a percentage of combined glucose, fatty acid and glutamine oxidation (n = 4 independent experiments). hj, Quantification of cellular metabolites from HEK293 cells (h, i) and MEFs (j). Metabolite abundance was normalized to those of WT cells (n = 5 independent experiments). Mean ± s.e.m.; one-way ANOVA with Dunnett’s multiple comparisons test (a, b); two tailed t-test (c, d, g); two-way ANOVA with Sidak’s multiple comparisons test (e).

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