This study investigates the role of lipid unsaturation in promoting BAX and BAK pore activity during apoptosis. By performing lipidomics analysis using lipid nanodiscs, the researchers found a significant enrichment of unsaturated lipids near BAX and BAK in apoptotic conditions. This enrichment was further confirmed in model membranes, isolated mitochondria, and cellular systems, with molecular dynamics simulations supporting these findings. The fatty acid desaturase FADS2 was identified as a key enzyme that enhances apoptosis sensitivity and the activation of the cGAS/STING pathway downstream of mtDNA release. The levels of FADS2 in lung and kidney cancer cells correlated with their sensitivity to apoptosis induced by co-treatment with unsaturated fatty acids. These results provide insights into how the local lipid environment affects BAX and BAK function during apoptosis, which may have implications for anticancer therapies.This study investigates the role of lipid unsaturation in promoting BAX and BAK pore activity during apoptosis. By performing lipidomics analysis using lipid nanodiscs, the researchers found a significant enrichment of unsaturated lipids near BAX and BAK in apoptotic conditions. This enrichment was further confirmed in model membranes, isolated mitochondria, and cellular systems, with molecular dynamics simulations supporting these findings. The fatty acid desaturase FADS2 was identified as a key enzyme that enhances apoptosis sensitivity and the activation of the cGAS/STING pathway downstream of mtDNA release. The levels of FADS2 in lung and kidney cancer cells correlated with their sensitivity to apoptosis induced by co-treatment with unsaturated fatty acids. These results provide insights into how the local lipid environment affects BAX and BAK function during apoptosis, which may have implications for anticancer therapies.