Fig. 6: Pilin-like proteins exclusively conserved among TANB77 order augments responsiveness of adaptive immunity through TLR4 signaling in antigen presenting cells.

a Diagram of a pilus gene cluster conserved in TANB77. The tree on the left represents the maximum likelihood phylogenetic tree of 129 representative MAGs of TANB77 species. The center plot illustrates the length and relative location of genes. The arrows represent coding sequences, and arrows with the same color indicate homologous sequences, while gray arrows represent non-conserved genes. The black lines represent contigs. The ‘x’ mark represents the end of the contig. The heatmap on the right provides information on the source of the genome and family. b A representative gene cluster derived from HRGM_Genome_2725, which is highlighted in red (a). The arrows represent coding sequences. The products of genes 927 to 932 are potentially involved in assembly machinery, and products of genes 932 to 938 are pilin-like proteins. Black triangles indicate the predicted promoters. c, d Mouse BMDCs (c) and human monocyte derived dendritic cells (d) were stimulated with indicated agonists targeting TLR2 and TLR4 and pilin 938 at indicated concentration for 24 hours. e After an hour of pre-treatment with TAK-242 inhibitors, BMDC were stimulated for 24 hours with TLR agonists and pilin 938 at the indicated concentration. f BMDC created from WT and TLR4 knockout (KO) bone marrow were stimulated with LPS and pilin 938 to see the TLR4 dependency of pilin 938 activation. g BMDCs were activated with either LPS or pilin 938 before being pulsed with GP33-41, then, these cells were co-cultured with P14 CD8⁺ T cells. c–g The in vitro experiments were conducted in triplet (n = 3) and the error bars represent mean ± SEM. The data are representative of two independent experiments except for (d) and (f) which were conducted once. The statistical analysis was conducted using One-Way ANOVA Tukey’s multiple comparisons test other than (e) and (f) where student t-test was used for analysis. Source data are provided as a Source Data file. (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).