Extended Data Fig. 6: Ferroptosis in genetically engeneered mice.
From: Ferroptosis of tumour neutrophils causes immune suppression in cancer
a. Ferroptosis related genes in PMN-MDSC from Fatp2flCre+ TB mice. qPCR of tumour PMN-MDSCs from Fatp2flCre− (WT) and Fatp2flCre+ (FATP2 KO) mice. N = 3. Mean ± SD are shown. P values shown on graphs were calculated in unpaired two-sided Student’s t-test. b. Oxidized PE in ALOX12/15 deficient mice PMN-MDSC. Content of oxygenated (PE(38:4+1[O]) PE(38:4+2[O]) PE(36:4+2[O]) and non-oxygenated (PE(38:4) PE species in PMN MDSC isolated from LLC TB Alox12/15flCre− and Alox12/15flCre− mice tumours. N = 3. Mean ± SD are shown. P values shown on graphs were calculated in unpaired two-sided Student’s t-test. c. Content of PE species containing oxygenated AA in PMN MDSC isolated from LLC TB WT and MPO KO mice. N = 3. d. Liproxstatin-1 inhibits MPO (0.05U)/H2O2(50 uM) induced formation of C18:0-Cl (left panel), PE-18:0p/20:4+3[O] (middle panel) and LPE(20:4+3[O]) (right panel) from PE(18:0p/20:4) in 20 mM phosphate buffer containing 100 mM NaCl and 100 uM DTPA, pH = 7.4, after 30 min incubation at 37 °C. The structure of PGE2 containing PE(18:0p/20+3[O]) was verified by three criteria (retention time (34 min), exact mass (m/z 798.5306 + 3.3 ppm) and MS/MS fragments (m/z 798→351, 271, 189, 113). LPE(20:4+3[O]) was identified as PGE2 containing species by exact mass (m/z 548.2636 + 3.4 ppm) and retention time (6 min). N = 4. Mean ± SD are shown. P values shown on graphs were calculated in unpaired two-sided Student’s t-test (a) or one-way ANOVA with correction for multiple comparsons.
