Supplementary Figure S3 from Neutrophils Mediate Protection Against Colitis and Carcinogenesis by Controlling Bacterial Invasion and IL22 Production by γδ T Cells

Figure S3. Neutrophils drive the expression of IL-23 by BMDMs. Related to Figure 3. A) Total number of ulcers in the colon of DSS-treated Csfr3+/+ (n=6) and Csf3r-/- (n=7) mice. B) IL-23 levels detected by ELISA in supernatants of BMDMs, neutrophils (NΦ) and BMDM-NΦ co-cultures after stimulation with GM-CSF+CpG. In indicated conditions neutrophils were pre-treated with diphenyleneiodonium (DPI) (10µM) or BMDM-NΦ were cultured in transwells (for all conditions n=6). C) Percentage of viable neutrophils (AnnexinV-/PI-) cultured alone or after co-culture with BMDMs. D) Schematic representation of the treatment schedule for anti-IL-22 treatment and neutrophil adoptive cell transfer during acute colitis. Days of treatment are indicated by red arrows. E) Percentage of body weight loss during DSS-induced acute colitis in Csf3r+/+(n=4), Csf3r-/-(n=4) and Csf3r-/- mice upon adoptive transfer of neutrophils treated with anti-IL-22 or isotype control (50µg/mouse) via i.p. injection (Csf3r-/- mice+Isotype n=3; Csf3r-/-+anti-IL-22 n=3). A) Representative data of five independent experiments. B) Representative data of three independent experiments. C) Representative of two independent experiments. E) One experiment. A) Unpaired Student’s t-Test. B-C) Multiple Student’s t-Test. E) Wilcoxon matched-pairs signed rank test. Data are mean ± SEM. *** p < 0.001 ** p < 0.01 * p < 0.05.