To generate Bax-KO cells, a pair of short guide-RNA (sgRNAs) were ordered from Integrated DNA Technologies (Supplementary Table 2). Oligonucleotides for guide RNAs were cloned into the pSpCas9(BB)-2A-GFP (PX458) plasmid (Addgene plasmid #48138) as described by Ran et al.73. BAX sgRNA plasmids were then transfected into DLD-1 cells using Lipofectamine®3000. 24–48 h after transfection, GFP positive cells were sorted and seeded individually into 96-well plates.

Knockdown of DUSP16 in C666-1 cells was performed by transfection with DUSP16 CRISPR/Cas9 KO Plasmid (sc-405727, Santa Cruz) and DUSP16 HDR plasmid (sc-405727-HDR, Santa Cruz) with UltraCruz® Transfection Reagent (sc-395739) according to the manufacturer’s instructions. Cells transfected with Control CRISPR/Cas9 plasmid (sc-418922) were used as control.

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