Microbiome transcriptional control of myd88 is dependent on Tlr2. a Venn diagram showing the overlap between the identities of 65 primarily suppressed transcripts displaying transcriptional sensitivity to all the different colonization modes tested in this study, and thus are strong markers of the shared transcriptional response, with 48 primarily induced transcripts displaying Tlr2 dependent transcriptional sensitivity to injection of the synthetic ligand Pam3CSK4. 11 transcripts were shared among the two sets, all of which are suppressed by colonization, but induced by Pam3CSK4 in a Tlr2 dependent manner. b Q-PCR analysis comparing the inflammatory il1b response to injection of Tlr2 ligand (PAM3CSK4), Tlr4 ligand (LPS) and control (PBS) in Tlr2-mut and Tlr2 + / + 1 h after injection. (mean ± s.e.m., n = 3 biological replicates, 10 embryos per group), *P ≤ 0.05 ***P ≤ 0.001; ****P ≤ 0.0001, by Student’s t-test. c Q-PCR analysis comparing the relative expression levels of myd88, fosl1a and cebpb, in conventionalized (CONVD) versus germ-free (GF) conditions in Tlr2 deficient mutants versus WT siblings. (mean ± s.e.m., n = 3 biological replicates, 15 embryos per group), *P ≤ 0.05; **P ≤ 0.01, by Student’s t-test. d Representative image of intestines excised from L-plastin stained embryos. Scalebar represents 100 µm. e The elevated leukocyte presence under germfree conditions in the WT siblings was not observed in Tlr2 mutants. Figure is representative of three independent replications, *P ≤ 0.05; by two-way ANOVA with Bonferroni correction for multiple comparisons, error bars represent standard error of the mean
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