Fig. 1.

Experimental design and functional validation of Hex-disrupted HL-60 mutants. (A) overview of human Hex genes, subunits and isoenzymes relevant to this study. (B) Experimental workflow for the (i) HL-60 CRISPR-Cas9 mutants and the (ii) Sandhoff disease and healthy donor neutrophils. †Repeat attempts to generate HEXA/HEXB double knockouts were unsuccessful as these mutants displayed severe growth retardation. (C) (i–ii), Determination of the total Hex and Hex A + S enzyme activity in the HL-60 cell lines using MUG(S) assays. Data are plotted as mean + SD, n = 3 technical replicates, ns, not significant, ^*P < 0.05, ^**P < 0.01, ^***P < 0.005, ^****P < 0.001 (HL-60 mutant vs wt).