Extended Data Fig. 4: Uba1 M41T mutation or partial inhibition of UBA1 with low-dose TAK-243 causes RIPK3 and Caspase-8 dependent inflammatory cell death.
From: Independent mechanisms of inflammation and myeloid bias in VEXAS syndrome
a, Cell death measured over 48 h after treatment with indicated stimuli in BMDMs of indicated genotypes 9 days after electroporation, quantified as percent YOYO-1+ cells among total cells. b, Cell death analysis after treatment with indicated stimuli in BMDMs of indicated genotypes, in presence or absence of 1 μM TAK-243, quantified as percent YOYO-1+ cells among total cells. c, Immunoblots of BMDMs of indicated genotypes 9 days after electroporation. d, Viable BMDM cell yields 9 days after electroporation relative to Uba1+/y (sgRosa26 electroporated) controls of matched Mlkl-Casp8 genotypes. Bars represent mean; error bars represent SEM. e, Cell death analysis of BMDMs of indicated genotypes after treatment with indicated stimuli in presence or absence of 100 nM TAK-243, quantified as percent YOYO-1+ cells among total cells. Lines in a, b, e represent mean values of triplicate wells, with 3 images taken per well for quantification; error bands represent SEM; results are representative of 2 independent experiments. f, g, Rectal temperatures and survival of C57BL/6 mice of indicated genotypes after i.v. injection of TNF (200 μg/kg) (f) or i.p. injection of LPS (5 mg/kg) (g), pre-treated 2 h prior with either vehicle or TAK-243 (3 mg/kg i.p.). Lines represent mean rectal temperatures for the indicated number of mice per condition. In g, experiments were performed 5 weeks after 10 Gy irradiation and bone marrow transplantation. P values comparing survival curves in (f-g) were calculated using the Log-rank (Mantel-Cox) test. Error bars in rectal temperature curves in (f-g) indicate SEM.
