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. 2020 May 11;23(6):101147. doi: 10.1016/j.isci.2020.101147

Figure 6.

Figure 6

Flagellin Treatment Does Not Enhance M Cell Differentiation in Enteroids

(A) IHC detection of CCL20 (red) in the FAE of Peyer's patches from PBS- and flagellin-treated aged mice. Nuclei detected with DAPI (blue). Scale bar, 20 μm. Broken line shows the apical FAE surface.

(B) Quantitation of the % area CCL20+ immunostaining in the FAE of Peyer's patches from mice in each group of (A). Each point is from an individual FAE. Horizontal line, median. n = 10–22/group from 3 to 4 mice. Statistical difference determined by Mann-Whitney.

(C–F) In vitro enteroids prepared from small intestinal crypts were treated with RANKL, flagellin or both. Expression of Ccl20 (C), Spib (D), Sox8 (E), and Gp2 (F) was compared by RT-qPCR 24 or 72 h later. Mean expression levels were normalized so that untreated enteroids at 24 h equaled 1.0. Data expressed as mean ± SEM. n = 3. Statistical differences determined by two-way ANOVA.

(G and H) Comparison of Tlr5 and Gp2 mRNA expression (G) in individual cell populations in deep CAGE sequence data from the FANTOM5 project of the FANTOM consortium (Forrest, 2014), and (H) in published mRNA-seq data from isolated GP2+ M cells (NCBI Gene Expression Omnibus: GSE108529 [Kimura et al., 2020]).