Figure 7. Distinct inflammatory cytokine induction by R. gnavus in lysozyme’s presence or absence.

(A) Experimental design for data in panels B-G. (B-C) Radar plots of cytokine/chemokine concentrations in the media of from WT and Lyz1^−/− MLN cells stimulated by supernatants from lysozyme-treated R. gnavus (red) versus untreated live R. gnavus culture (green). (D-E) Representative dot blots and summary quantification from two independent experiments (each in 2 replicates). (F) Western blots of pStat6 and pStat3 in lysates of WT and Lyz1^−/− MLN cells treated analogous to B-E. (G) Experimental design for data in panels H-K. (H-J) Bulk RNAseq of ileal mucosa 14-days after R. gnavus gavage (n=3). Increased tuft cell signature, type 2 immune response, and IL-13 response in R. gnavus colonized mice in GSEA analysis. (K) Differential expression of IL-13 responsive genes in R. gnavus-colonized Lyz1^−/− compared to PBS-gavaged Lyz1^−/− mice. (L) Experimental design for data in panels M-O showing that DSS colitis was induced in R. gnavus colonized WT and Lyz1^−/− littermates following Abx treatment. (M) Body weight changes during DSS exposure and recovery in Abx-precleared WT and Lyz1^−/− mice with or without R. gnavus association (n number indicated; 2 independent experiments). (N) Colitis activity scores in the colon of DSS-treated Lyz1^−/− mice with or withour R. gnavus association. (O) Representative H. & E. images of DSS colons of Abx-treated and R. gnavus colonized WT and Lyz1^−/− mice. All bar graphs display mean ± SEM. See also Figure S7.