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. 2024 Jun 19;52(14):8271–8285. doi: 10.1093/nar/gkae519

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© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Simultaneous loss of redundant formaldehyde-detoxification pathways causes embryonic lethality and developmental delay, which is enhanced in NER-deficient animals. (A) Phylogenetic tree of C. elegans proteins showing similarity to human ALDH2. Numbers in red branch support values. (B) Experimental scheme (see Materials and methods for details). Cartoons were created with BioRender.com. (C) Heatmap combining data of development and embryonic lethality of animals exposed to alh-1(RNAi). The displayed values are the mean percentages of events counted on the plates (stages and dead embryos) after 48 h of development for three technical replicates. The significance was determined via the two-way ANOVA, followed by the Tukey's multiple comparison test, for which counts *P < 0.05, **P < 0.01, ***P < 0.001, ****P< 0.0001. A full statistical analysis between all the groups is available in the supplementary tables. (D) Percentages of L1/L2 stage animals for the various mutant backgrounds with alh-1(RNAi). (E) Percentages of dead embryos counted on plates. The significance in D and E was determined using a one-way ANOVA followed by the Tukey's multiple comparison test reporting adjusted p values for each comparison. (F) Viability plots for Epstein-Barr virus (EBV) immortalized lymphoblastic cells from patients with mutations in the genes depicted in the scheme growth. Cells were exposed to formaldehyde (FA) in presence/absence of a cocktail containing the ADH5 inhibitor N6022 (10 μM) and the ALDH2 inhibitor Disulfiram (0.1 μM) (n = 3). (G) Table showing the IC50 for FA calculated from the data in F using GraphPad.