Extended Data Fig. 8: Loss of endothelial Pten results in increased polyamine levels.
(a) Schematic representation of control and PteniΔEC eWAT explant experiment. eWAT from 5 weeks of age control and PteniΔEC mice were cut in ~25 mg pieces, placed in 500ul of DMEM low glucose + 0,5% FFA free BSA, and incubated for the indicated time points. (b) Experimental design of conditioned media collection and fractioning. A 3 kDa cut off filter retained the protein and vesicle fraction, whereas the metabolite-rich fraction was collected as the flow-through. The lipolytic effect of both fractions was tested on eWAT explants of wild-type mice. (c) Label-free 13C fraction of metabolites related to polyamines biosynthesis in adipose-ECs from control and PteniΔEC mice (n = 6 per group) from Fig. 6g. MET: Methionine; SAM: S-Adenosylmethionine; dcSAM: Decarboxylated S-adenosylmethionine; MTA: 5´ Methylthioadenosine. Data represent mean ± SEM (error bars). Statistical analysis was performed by t-test (Extended Data Fig. 8c).
