Extended Data Fig. 7: Effects of Uba1 mutation on HSPCs and DAMPs.

a, Immunoblots of Uba1^+/y or Uba1^M41T/y HSPCs 5 days after electroporation. b, Frequency of indicated cell populations (L: Lineage⁻, S: Sca-1⁺, K: c-KIT⁺, Gr-1: Ly-6G and/or Ly-6C) among HSPCs 6 days after electroporation with ABE8e-SpRY plus sgGFP (Uba1^+/y) or sgM41T (Uba1^M41T/y) (n = 5 biological replicates per genotype). ****P < 0.0001. c, Representative flow cytometry plot of CD11b and F4/80 expression 6 days after electroporation of Uba1^+/y or Uba1^M41T/y HSPCs, % of cells in each quadrant is indicated. d, Wright-Giemsa stain of Uba1^+/y or Uba1^M41T/y HSPCs 8 days after electroporation. e, Immunoblots of Uba1^+/y or Uba1^M41T/y HSPCs 3 days after electroporation. f, Cell death measured over 24 h in HSPCs of indicated genotypes treated with indicated stimuli 4 days after electroporation. Lines represent mean values of triplicate wells, with 3 images taken per well for quantification; error bands represent SEM. g, Analysis of cell cycle (% of cells in S2, G, or M phase) by EdU incorporation vs Propidium iodide stain, viability, and M41T frequency (via Sanger sequencing) at the indicated timepoints after electroporation of HSPCs of the indicated genotypes (n = 4 biological replicates). h, IL-1α (n = 3), IL-1β (n = 3) and IL-18 (n = 6) release by Uba1^+/y or Uba1^M41T/y BMDMs treated with indicated stimuli for 24 h. i, j Representative flow cytometry plot (i) and quantification (j) of myeloid differentiation of HSPCs 96 h after treatment with IL-1α or IL-1β (n = 2 per condition). Dots in b, g, h, j represent biological replicates; bars represent mean values; error bars represent SEM. Results are representative of 3 (a-d) or 2 (e-j) independent experiments. P value for % M41T graph (right) in panel g was calculated using paired two-sided t-test. Otherwise, P values in b, g, h were calculated using unpaired, two-sided t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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