Fig 2: Relationship between phosphorylation of intracellular signal kinases and lysophosphatidylcholine (lysoPC) effects through long‐chain acyl‐coenzyme A synthases 5 (ACSL5). Levels of phosphorylated (p‐) phosphoinositide 3‐kinase (PI3K), Akt, mTOR, GSK‐3β, TSC2, Raf, MEK, ERK, RSK and MSK proteins were measured in SPC‐A1 cells 6 h after treatment with lysoPC. Regulatory effects of PI3K and ERK in reactive oxygen species (ROS) production quantified by DCF fluorescence intensity (B), ACSL5 gene expression (C) and cell proliferation rate (D) were assayed in SPC‐A1 cells pretreated with PI3K inhibitor LY294002 or ERK inhibitor GDC0994 for 2 h and followed by lysoPC treatment for 6 h (n = 6) or in ACSL5 NC or ACSL5 KD cells pretreated with inhibitors of PI3K and ERK for 2 h (n = 6). Protein levels of ACC and FASN in SPC‐A1 cells pretreated with vehicle or PI3K/ERK inhibitors were measured 6 h after lysoPC treatment (E). * and ** or # and ## stand for the p‐values less than .05 and .01, respectively, as compared with cells treated with vehicle or lysoPC

Fig 3: Validation of long‐chain acyl‐coenzyme A synthases 5 (ACSL5)‐dependent roles after down‐regulation of ACSL5. Transcriptomic profiles of ACSL5 NC and ACSL5 KD with vehicle or lysophosphatidylcholine (lysoPC) were measured by bulk RNA‐seq, of which the heatmap of top 50 lipid‐associated differentially expressed genes (A), the exact levels of ACSL family member genes (B) and the signal pathway enrichment of those genes (C) were presented (n = 3 per group and repeated thrice). ACSL5‐dominated lipid metabolism gene networks using Cytoscape (D), heatmap of genes involved in fatty acid biosynthesis, degradation and elongation (E) and volcano plot of lipid‐related genes (F) between ACSL5 NC and ACSL5 KD cells treated with vehicle or lysoPC were calculated, on basis of up‐regulated genes with fold changes > 1.5 and down‐regulated genes with fold changes < 0.67 (p < .05). * and ** or # and ## stand for the p‐values less than .05 and .01, respectively, as compared with ACSL5 NC cells treated with vehicle or lysoPC. FASN: fatty acid synthase; OXSM: 3‐Oxoacyl‐ACP Synthase, mitochondrial; ACSL5: acyl‐CoA synthetase long‐chain family member 5; ACAT1: acetyl‐CoA acetyltransferase 1; ALDH9A1: aldehyde dehydrogenase 9 family member A1; ACADM: acyl‐CoA dehydrogenase medium chain; ECI2: enoyl‐CoA delta isomerase 2; HADHA: hydroxyacyl‐CoA dehydrogenase trifunctional multienzyme complex subunit alpha; HADHB: hydroxyacyl‐CoA dehydrogenase trifunctional multienzyme complex subunit beta; ELOVL4: ELOVL fatty acid elongase 4; PPT1: palmitoyl‐protein thioesterase 1; ACOT7: acyl‐CoA thioesterase 7; and ELOVL3: ELOVL fatty acid elongase 3