Extended Data Fig. 1: Naïve ESCs grow without exogenous leucine.
From: Amino acid intake strategies define pluripotent cell states
(a) Population doublings of S/L- and S/L + 2i-cultured ESCs subjected to glutamine (Gln) replete or depleted conditions for 48 h. (b) Relative viability (measured by PI exclusion) of S/L- and S/L + 2i-cultured ESCs subjected to leucine (Leu) deprivation for 48 h. Viability is expressed relative to leucine-replete controls. (c) Teratoma formation from S/L + 2i-cultured ESCs subjected to leucine (Leu) replete or depleted conditions for 24 h. Each injection resulted in teratoma formation within two weeks (6 per group); two tumors per group were subjected to histology analysis with similar findings. Representative images of hematoxylin and eosin stained tumor tissue showing differentiation into endoderm, mesoderm and ectoderm-derived tissues. (d) Population doublings of S/L- and S/L + 2i-cultured ESCs (ESC-2) subjected to leucine (Leu) depleted conditions for 48 h. (e) Population doublings of S/L- and S/L + 2i-cultured ESCs subjected to isoleucine (Ile), phenylalanine (Phe), or valine (Val) depleted conditions for 48 h. (f) Cell volume of S/L- and S/L + 2i-cultured ESCs subjected to leucine (Leu) replete or depleted conditions for 48 h. (g) Change in total cellular volume of S/L- and S/L + 2i-cultured ESCs (ESC-1 and ESC-2) subjected to leucine (Leu) depleted conditions for 48 h. (h) Change in protein content of S/L- and S/L + 2i-cultured ESCs subjected to leucine (Leu) depleted conditions for the indicated times. Data are mean ± SD, n = 3 independent samples. Significance was assessed by unpaired two-tailed t test (b,d-g) and two-way ANOVA with Sidak’s multiple comparisons post-test (h). Scale bar, 50 µm.
