Article Figures & Data
Figures
- FIG 1
Outline of our method of identifying uncultured rumen bacteria that take up glucose with 2-NBDG.
- FIG 2
Microscopic image of mixed rumen bacteria incubated in 2-NBDG. Images show cells before cell sorting (A, C, and E) and after two sequential rounds of sorting (C, D, and F). Cells labeled with 2-NBDG are green. Images are from one of two independent experiments.
- FIG 3
Flow cytometry histogram showing bacteria labeled with 2-NBDG and their separation by fluorescence activated cell sorting (FACS). Shown are cells before sorting (A), after one round of sorting (B), and after two rounds (C). Over 95% purity was achieved. Panels B and C present results of a postsort analysis. SYTO 62 was added to stain DNA. Histograms are from 1 of 15 independent experiments.
- FIG 4
Violin plot of the 10 most abundant bacteria that take up 2-NBDG. A single-sided Wilcoxon signed-rank test was performed to test if abundance was >0 after accounting for impurity from unsorted samples. Each observation represents 1 of 15 independent experiments. See Fig. S3 for the full list of bacteria.
- FIG 5
Microscope image that shows absence of cross-feeding of 2-NBDG between Streptococcus equinus JB1 (coccus) and Anaerovibrio lipolyticus 5S (crescent). Cocultures of S. equinus JB1 and A. lipolyticus 5S were given 2-NBDG (5 mM). S. equinus JB1 takes up 2-NBDG and is labeled green, whereas A. lipolyticus 5S remains unlabeled; the fluorescent group in 2-NBDG is not transferred from the former to the latter. Images are from one of two independent experiments.
Tables
Supplemental material
- Supplemental file 1 -
Flow cytometry histogram showing that 2-NBDG is not taken up by dead cells (Fig. S1); analysis of α- and β-diversity of samples (Fig. S2); heat map for relative abundance of bacteria that are present in FACS-sorted cells and take up 2-NBDG (Fig. S3); violin plot of ten most abundant bacteria identified by anaerobic culture using RGA or RA medium (Fig. S4); flow cytometry histogram showing stability of 2-NBDG label in bacterial cells under aerobic conditions (Fig. S5); analysis of α- and β-diversity for two different DNA extraction methods (Fig. S6).
PDF, 1.2M
- Supplemental file 2 -
Percentages of rumen bacteria gated as positive for 2-NBDG during cell sorting (Data Set S1); relative abundances of amplicon sequence variants in all sample types (Data Set S2); sequences and assigned taxonomy of amplicon sequence variants (Data Set S3); matches between previously cultured strains and amplicon sequence variants that take up 2-NBDG (Data Set S4).
XLSX, 1.6M
- Supplemental file 1 -
