Extended Data Fig. 3: INSeq analysis of relative nutritional requirements for A. muciniphila to grow in mucin medium and the role of putative glycan hydrolases.
Plot of INSeq data from Tn mutant pools grown for eight generations in mucin medium where each dot represents all inserts in a specific gene. Genes that belong to KEGG amino acid biosynthesis pathways are highlighted for cultures grown in (a) mucin medium and (b) mucin medium supplemented with Phytone. Predicted glycosyl hydrolases for A. muciniphila BAA-835 were identified using the CAZy database and highlighted on the INSeq plot for cultures grown in (c) mucin and in (d) mucin medium with Phytone. Statistical analysis on INSeq data was performed with a Mann-Whitney Utest. (e) Droplet-seq analysis of A. muciniphila grown in mucin medium microdroplets. Tn mutants (Arrayed Pool) were injected into a microfluidic device at a low density to generate on average less than one bacterium per droplet. The graph displays the INSeq analysis and Log2 fold change for cultures grown in mucin in batch culture (8 generations) versus single cell growth in droplets (72 h). Selected genes that were depleted in one condition relative to the other are highlighted on the plot. GH, glycosyl hydrolase.
