Supplementary Figure 5: Intestinal IgA production is normal in Il17f^−/− mice.

(a, b) Proportion of IgA⁺ B cells in the colon of indicated mice were examined by flow cytometry (n=3–6/group). (a) Cells are pre-gated on cLP 7AAD^-Lymphocytes (left) or 7AAD^-CD19⁺Lymphocytes (right) and numbers in dot plot panels indicate percentages of IgA⁺ B cells in total cLPLs (left) or total cLP B cells (right). (b) Percentages of IgA⁺ B cells in cLP total B cells in indicated mice are shown (WT n=4, l17f^–/– n=3, Il17a^–/– n=5, l17f^–/–Il17a^–/– n=3). WT vs Il17a^–/– *P=0.0158, Il17f^–/– vs Il17a^–/–Il17f^–/– *P=0.0356. (c) Expression of Pigr in the colon of indicated mice were examined by qPCR (WT n=12, l17f^–/– n=8, Il17a^–/– 7). ***P=0.0006. (d-f) Fecal microflora were harvested from WT, Il17f^−/− or Il17a^−/− mice and were stained by anti-IgA Ab. The IgA⁺ or IgA^- commensal bacteria were purified by FACS sorting, and were further identified as IgA-specific/nonspecific commensals. (d) IgA-bound/non-bound bacterial population before and after the purification was confirmed by flow cytometry. (e) Indicated IgA-specific/nonspecific commensals in indicated groups of mice are shown by the ratio of % in purified total IgA⁺ bacteria / % in purified total IgA^- bacteria. (f) IgA-specific/nonspecific commensals in indicated individual mouse are shown in heat map by using the same data described in (e) (n=3/group). Data in (a, d) are representative of two independent experiments with similar results. Data in (b, c, e) are expressed as means ± SD (for b-c, one-way ANOVA followed by Tukey’s multiple-comparisons test).