Extended Data Fig. 2: Transcriptional and functional changes in UBA1^mut cells in vitro.

a, Schematic representation of the editing procedure and in vitro experimental design in male human HSPCs. b-c, Western blot analysis of UBA1a, UBA1b, UBA1c protein expression (b), poly-ubiquitylated proteins (Poly-Ub) and BiP expression level (c) in UBA1^wt and UBA1^mut HSPCs. β-actin was used as protein loading control (n = 2 biological replicates from different HSPC donors). d, Volcano plots showing fold changes of up- (red) and down- (green) regulated genes comparing UBA1^mut and UBA1^wt HSPCs one (left) or seven (right) days after editing. Non-significant genes are shown in grey. Wilcoxon rank-sum test with Benjamini-Hochberg correction. e, Percentage of male primary T cells carrying the UBA1 edit over time after editing (n = 6 biological replicates). Median with IQR. Wilcoxon matched-pairs signed rank test. f, Percentage of live T cells at the endpoint of the experiment in ‘e’ (n = 6 biological replicates). Median with IQR. Wilcoxon matched-pairs signed rank test. g, Western blot analysis of cleaved caspase-3 in UBA1^wt and UBA1^mut T cells. β-actin was used as protein loading control. Panel a created with BioRender.com.