![Temporal changes in neutrophils during infection. Mice were inoculated intraperitoneally (IP) with E coli (105 CFU, self-limited dose; see “Methods”). Exudates and BM cells were collected at 0, 12, 24, 48, and 72 hours after inoculation. Numbers of exudate (A); whole blood (B); BM neutrophils (C); and BM granulocyte progenitors, LSKs and GMPs (D), were determined by flow cytometry. (E) Screen captures of RvD1 from targeted LC-MS/MS analysis. (Top) BM RvD1–targeted scheduled multiple reaction monitoring (MRM) in negative-ion mode for mass-to-charge (m/z) 375 → 215 (“→” indicates fragmentation/transition). (Bottom) Synthetic RvD1. MRM signal-to-noise ratio of >80. (F) Screen captures of RvD4 from targeted LC-MS/MS analysis. (Top) BM RvD4–targeted scheduled MRM in negative-ion mode for m/z 375 → 101. (Bottom) Synthetic RvD4. MRM signal-to-noise ratio of >40. (G) Screen capture of BM RvD4 MS/MS fragmentation. Inset: RvD4 structure and proposed fragmentation. BM RvD4 MS/MS fragmentation spectrum matched to an unbiased library of synthetic RvD4 standard with a fit score of 94.5%. BM-RvD4 MS/MS spectral ions gave a parent ion mass of m/z 375 (M-H) and daughter ions of m/z 357 (M-H–H2O), 339 (M-H–2H2O), 313 (M-H–H2O-CO2), 295 (M-H–2H2O-CO2), 255 (273–H2O), 233 (277–CO2), and 101 (M-H–CHOH-[CH]6-CH2-[CH]4-CHOH-CH2-[CH]2-CH2-CH3). (E-F) Screen captures taken from Sciex OS version 1.7.036606 (Explorer Mode). Note the additional digits in the screen captures in panel G are the default setting (the number [amu] in the second decimal place does not accurately reflect the mass of the ions).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/142/6/10.1182_blood.2022019145/2/blood_bld-2022-019145-gr1.jpeg?Expires=1738071108&Signature=WoqIzocfADJBTpngn4xtxTSbxqrKfeO0eU0~3gox~QQ-QnCWQUN-1ssiAUt1CbsftkpmiJL6CmsXkE8savzgqzYXETWzQWkY9EHHmJ6moZrFk2yCFiRTi5f6ubtfZtsuMWc28bxXzA8T-VLqAMTohITaq03l~auBGXhXFtAkTCUSEsC7Cr3NYakXOBkiqTo4jr7VneBQ8Ax0j39oChtP9A9U3hy5H1uNFBL1~vjlkS~~JnBOCruUbkwiNNWXje5Pp72qe5rN50tMT1RiXExuScbwcuU-6vaFFLAraVIELBJ0fijmVD27UWjN95NfmxmqFZxp7-HOmOq72R9dGmzfUg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Temporal changes in neutrophils during infection. Mice were inoculated intraperitoneally (IP) with E coli (105 CFU, self-limited dose; see “Methods”). Exudates and BM cells were collected at 0, 12, 24, 48, and 72 hours after inoculation. Numbers of exudate (A); whole blood (B); BM neutrophils (C); and BM granulocyte progenitors, LSKs and GMPs (D), were determined by flow cytometry. (E) Screen captures of RvD1 from targeted LC-MS/MS analysis. (Top) BM RvD1–targeted scheduled multiple reaction monitoring (MRM) in negative-ion mode for mass-to-charge (m/z) 375 → 215 (“→” indicates fragmentation/transition). (Bottom) Synthetic RvD1. MRM signal-to-noise ratio of >80. (F) Screen captures of RvD4 from targeted LC-MS/MS analysis. (Top) BM RvD4–targeted scheduled MRM in negative-ion mode for m/z 375 → 101. (Bottom) Synthetic RvD4. MRM signal-to-noise ratio of >40. (G) Screen capture of BM RvD4 MS/MS fragmentation. Inset: RvD4 structure and proposed fragmentation. BM RvD4 MS/MS fragmentation spectrum matched to an unbiased library of synthetic RvD4 standard with a fit score of 94.5%. BM-RvD4 MS/MS spectral ions gave a parent ion mass of m/z 375 (M-H) and daughter ions of m/z 357 (M-H–H2O), 339 (M-H–2H2O), 313 (M-H–H2O-CO2), 295 (M-H–2H2O-CO2), 255 (273–H2O), 233 (277–CO2), and 101 (M-H–CHOH-[CH]6-CH2-[CH]4-CHOH-CH2-[CH]2-CH2-CH3). (E-F) Screen captures taken from Sciex OS version 1.7.036606 (Explorer Mode). Note the additional digits in the screen captures in panel G are the default setting (the number [amu] in the second decimal place does not accurately reflect the mass of the ions).
