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Fig. 2 | Microbiome

Fig. 2

From: Gut microbiota–derived metabolite 3-idoleacetic acid together with LPS induces IL-35+ B cell generation

Fig. 2

Reg 4 promotes the accumulation of IL-35+ cells in adipose tissues. a Flow cytometry of p35+CD19+, p35+CD4+, p35+Ebi3+CD19+, and p35+Ebi3+CD4+ cells in the fat pat adipose tissues of Reg4 KO (R4KO) and control WT (WT1) mice and in huREG4IECtg (HuR4) mice and their littermate controls (WT2) fed a HFD for 3 months. b Flow cytometry of p35+CD19+, p35+CD4+, p35+Ebi3+CD19+, and p35+Ebi3+CD4+ cells in the subcutaneous adipose tissues of Reg4 KO (R4KO) and control WT (WT1) and in huREG4IECtg (HuR4) mice and their littermate controls (WT2) fed a HFD for 3 months. c Flow cytometry of CD19+IgM+ and CD19+IgD+ cells in the subcutaneous adipose tissues of huREG4IECtg (HuR4) mice and their littermate controls (WT2) fed a HFD for 3 months. d Flow cytometry of IgD+p35+ cells in the adipose tissues of huREG4IECtg (HuR4) and control littermate WT mice (WT2) fed a HFD for 3 months. e Immunostaining of IgD+p35+ in the adipose tissues of huREG4IECtg (HuR4) and control littermate WT mice (WT2) fed a HFD for 3 months. One representative result is shown from each group. Iso. ctr Ab, isotypic antibody. f qRT-PCR of IL-35 subunits (Ebi3 and p35), TGFβ and IL-10 in the adipose tissues of Reg4 KO (R4KO), WT (WT1) and huREG4IECtg (HuR4) mice and their control WT littermates (WT2) fed a HFD for 3 months (mixed sample). g ELISA of IL-35 in Reg4 KO (R4KO), WT (WT1) and huREG4IECtg (HuR4), WT (WT2) mice with (HFD) or without (normal) HFD feeding for 3 months. The data in a, b, c, d, and f were from three independent experiments; the data in g were from one representative experiment. Scale bars = 40 μm; Student’s t test in all panels, mean ± SD; *p < 0.05, **p < 0.01, and ***p < 0.001; NS, no significance

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