Fig. 1: ACSL3 drives prostaglandin synthesis in NSCLC.
From: The ACSL3-LPIAT1 signaling drives prostaglandin synthesis in non-small cell lung cancer
a PC, PS, PE, and PI (mol%/sample) lipid alterations 72 h after ACSL3 knockdown in A549 cells. Cells were transduced with either an empty vector control (pLKO) or an shRNA against ACSL3 (shACSL3 #1), and 72 h later lipids were extracted and analyzed by mass spectrometry-based shotgun lipidomics. PC phosphatidylcholine, PS phosphatidylserine, PE phosphatidylethanolamine, PI phosphatidylinositol. n = 4/group. Data are presented as mean ± SD. C38:4-PI4P (b), C38:4-PI(4,5)P2 (c) peak areas (normalized to internal standard and protein content) after ACSL3 knockdown in A549 cells. Cells were transduced with either an empty vector control (pLKO) or an shRNA against ACSL3 (shACSL3 #1), and 72 h later lipids were extracted and analyzed by ultra-performance liquid chromatography-tandem mass spectrometry. n = 4/group. Data are presented as mean ± SD. d Immunoblot analysis of ACSL3 in A549 and A427, H1264, and H358 NSCLC cell lines transduced with either an empty vector control (pLKO) or two different shRNAs against ACSL3 and extracted 72 h later. e PGE2 ELISA assay for the indicated cell lines. Cells were transduced with either an empty vector control (pLKO) or two different shRNAs against ACSL3. PGE2 production was analyzed 24 h later. n = 3/group. Data are presented as mean ± SD. f Quantification of prostaglandin E2, D2, and I2 from p53flox/flox;Acsl3+/+ and p53flox/flox;Acsl3–/– lungs, or from KrasG12D/WT;p53flox/flox;Acsl3+/+ and KrasG12D/WT;p53flox/flox;Acsl3–/– macrodissected tumors, 10 weeks after tumor onset. Samples were extracted and analyzed by ultra-high-performance liquid chromatography with tandem mass spectrometry n = 3/group. Data are presented as mean ± SEM. Statistical analyses were done using two-tailed unpaired Student’s t test or one-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
