Extended Data Fig. 1: Spermidine is decreased in NAT1-deficient cells.
From: Spermidine mediates acetylhypusination of RIPK1 to suppress diabetes onset and progression
(a) Western blotting analysis of Nat1 knockout and WT MEFs. (b) Quantification of the relative levels of isoniazid (1 mM, 4 hours) and acetylated isoniazid in wild-type and Nat1 knockout MEFs using HPLC-MS. N = 6 in each group. P values were determined using two-tailed Student’s t-test. Data are presented as means ± SEM. (c) WT and Nat1 knockout MEFs pretreated with polyamine oxidase inhibitor MDL72527 (20 μM) for 1 hour were treated with TNFɑ (10 ng/ml) and SM164 (50 nM) to induce RDA with or without putrescine (50 or 100 μM) pretreatment for 4 hours or Nec-1s (10 μM) pretreatment for 1 hour for indicated periods of time and cell death was measured by SytoxGreen positivity. N = 3 in each group. P values were determined using two-way ANOVA post hoc Bonferroni’s test. Data are presented as means ± SEM. (d) WT and Nat1 knockout MEFs were treated with TNFɑ (10 ng/ml)/SM164 (50 nM)/zVAD-fmk (25 μM) to induce necroptosis with or without putrescine (50 or 100 μM) pretreatment for 4 hours or Nec-1s (10 μM) pretreatment for 1 hour for indicated periods of time and cell death was measured by SytoxGreen positivity. N = 3 in each group. P values were determined using two-way ANOVA post hoc Bonferroni’s test. Data are presented as means ± SEM.
