Fig. 6

IL-35 promotes resistance to HFD-induced obesity. a Study design of the in vivo mouse experiment. b Changes in the body weights of mice fed a HFD. Mice were fed a HFD for 10 weeks and then transplanted with IL-35⁺ B cells (+IL-35 cells) treated with (+IL-35 siRNA) or without siRNA. Other cells, isolated B cells from spleen. c Flow cytometry of F4/80⁺CD11C⁺, F4/80⁺CD206⁺ cells, IFNγ⁺CD4⁺, Foxp3⁺CD4⁺ in the fat pads of WT1 and Reg4 KO (R4KO) mice and WT2 and huREG4^IECtg (HuR4) mice fed a HFD and then transplanted with IL-35⁺ B cells (+IL-35 cells) treated with (+IL-35 siRNA) or without siRNA (n = 3). Other cells, isolated B cells from spleen. d Study design of the in vivo mouse experiment. e Changes in the fat pad weights of mice fed a HFD for 10 weeks and treated with or without rIL-35 and IL-35 neutralizing antibodies. The mice were fed a HFD for 10 weeks and then injected with rIL-35 or IL-35 neutralizing antibodies via their inguinal fat pad tissues. WT (WT1) and Reg4KO (R4KO) mice treated with PBS (NC) or rIL-35 (+IL-35); WT (WT2) and huREG4^IECtg (HuR4) mice treated with an isotype antibody (+Iso. Ab) or a IL-35 neutralizing antibody (+IL-35 Ab) (n = 7, per group). f, g Flow cytometry of F4/80⁺CD11C⁺, F4/80⁺CD206⁺, IFNγ⁺CD4⁺, and Foxp3⁺CD4⁺ cells in the fat pad tissues of WT (WT1) and Reg4 KO mice (R4KO) treated with or without rIL-35 in (f) and in WT (WT2) and huREG4^IECtg mice (HuR4) treated with or without an anti-IL-35 neutralizing antibody in (g) (n = 3). h ELISA of IL-35 in the fat pad tissues of mice injected with rIL-35- or IL-35-blocking antibodies. The data in b, c, f, and g were from three independent experiments; the data in e and h were from one representative experiment. Analysis of variance in b; Student’s t test in the other panels, mean ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001