Fig. 2: A secondary knockdown screen identifies EXO1 as a formaldehyde resistance gene. | Nature Communications

Fig. 2: A secondary knockdown screen identifies EXO1 as a formaldehyde resistance gene.

From: A CRISPR-Cas9 screen identifies EXO1 as a formaldehyde resistance gene

Fig. 2

a Left. 20 drop-out hits were selected by their normZ score (lower than −3 on both day 9 and day 15, and a FDR lower than 0.05). Right. The 20 drop-out hits are presented by STRING. Green: formaldehyde catabolic processes, blue: FA pathway genes, yellow: transcription-coupled nucleotide excision repair genes, red: EXO1, gray: other genes non-assigned to any specific pathway. b Cell survival in RPE-1 cells transfected with siCTL or individual siRNAs targeting the 20 drop-out hits from the CRISPR-Cas9 screen. Data are presented with ±SEM from three independent experiments. *p < 0.1, ***p  <  0.001 and ****p  <  0.0001 (ordinary one-way ANOVA). c Schematic overview of domains in the human EXO1 protein. d Survival curve of RPE-1 EXO1 wild-type (WT) cells and RPE-1 EXO1 knockout clones (KO 7, KO 11) treated with different concentrations of formaldehyde for 96 h. Data are presented with ±SEM from three independent experiments. Protein level of EXO1 in RPE-1 WT, KO 7, KO 11 cells, with vinculin as loading control. e Survival curve of RPE-1 EXO1 WT with AAVS1 empty vector (WT + EV) cells, KO 11 with AAVS1 empty vector (KO 11 + EV) cells and KO 11 complemented with WT EXO1 (KO 11 + WT EXO1) cells treated with different concentrations of formaldehyde for 96 h. Data are presented with ±SEM from three independent experiments. Protein level of EXO1 in RPE-1 AAVS1 complemented cells, with α-tubulin as loading control. f Scheme for generating single copy of EXO1 WT complemented cells using integration at the AAVS1 single locus.

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