<i>Bacillus anthracis</i> and <i>B</i>. <i>cereus</i> strains used in this study.

<p><i>Bacillus anthracis</i> and <i>B</i>. <i>cereus</i> strains used in this study.</p

Reactivity of monoclonal antiserum EAII-6G6-2-3, specific for <i>Ba</i> neutral cell wall polysaccharide [19], with HF-SCWPs from <i>Bc</i> group strains.

<p>Meso Scale Discovery (MSD) multi-array high bind 96 well plates were coated with a fixed concentration of HF-SCWP antigen (2 μg/ml) from different <i>Bacillus</i> species and probed with the monoclonal antibody in serial two fold dilutions. Bound antibody was detected by using 2.5 μg/ml of sulfo-tagged goat anti-mouse IgM detection antibody. Data points are the average of three independent experiments. Error bars represent one standard error. Reactivity reported as effective concentration (EC₅₀) titer in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0183115#pone.0183115.t004" target="_blank">Table 4</a></b>.</p

Expanded region of proton NMR spectra identifying the locations and area ratios of the Gal substituents on HF-SCWPs from <i>B</i>. <i>cereus</i> strains.

<p><b>(A)</b> human isolate <i>B</i>. <i>cereus</i> G9241; <b>(B)</b> great ape isolate <i>B</i>. <i>cereus</i> CA (Cameroon); <b>(C)</b> great ape isolate <i>Bc</i> CI (Côte d’Ivoire). The α-anomeric signal originates from the free reducing end of all these polysaccharides, and co-migrates with a new residue (<b>K</b>) in the <i>Bc</i> CA and <i>Bc</i> CI SCWPs. This reducing end (α-GlcNAc residue) was removed by borodeuteride reduction for subsequent experiments, to facilitate integration and characterization of the residue <b>K</b> system. The percentage of Gal (<b>G</b>) and Gal-Gal disaccharide (<b>J</b>+<b>K</b>) substitution at ManNAc residues (<b>B</b>+<b>B′</b>+<b>B″</b>) is estimated by examination of the signal areas for these residues. The residue <b>G</b> spin system undergoes a shift at several positions, and is designated residue <b>J</b>, when it is substituted by residue <b>K</b> (refer to <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0183115#pone.0183115.t002" target="_blank">Table 2</a></b>).</p

600-MHz ¹H and ¹³C NMR parameters observed for the HF-released secondary cell wall polysaccharide released by HF treatment of great ape isolate <i>B</i>. <i>cereus</i> strain CA cell walls^a.

<p>600-MHz ¹H and ¹³C NMR parameters observed for the HF-released secondary cell wall polysaccharide released by HF treatment of great ape isolate <i>B</i>. <i>cereus</i> strain CA cell walls<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0183115#t002fn003" target="_blank">^a</a>.</p

Percent and type of structural substituent at the 3-position of backbone ManNAc residues in HF-SCWPs from examined infection-associated <i>B</i>. <i>cereus</i> strains.

<p>Percent and type of structural substituent at the 3-position of backbone ManNAc residues in HF-SCWPs from examined infection-associated <i>B</i>. <i>cereus</i> strains.</p

Immunochemical analysis of HF-SCWP with monoclonal and polyclonal antisera.

<p>Immunochemical analysis of HF-SCWP with monoclonal and polyclonal antisera.</p

Proton NMR spectra comparing the <i>Bc</i> Cameroon HF-SCWP before and after reduction of the reducing end with borodeuteride.

<p>The effect of borodeuteride reduction on the signal at (<b>K</b> + α) is demonstrated: <b>(A)</b> native <i>Bc</i> CA HF-SCWP before reduction; <b>(B)</b> the <i>Bc</i> CA HF-SCWP after reduction of the reducing end (red-HF-SCWP). The contribution from the α-reducing end anomeric signal to residue <b>K</b> signal area was almost completely eliminated by 1 h treatment, allowing unambiguous assignment of residue <b>K</b> system and area measurements. Following reduction, the <b>K</b>/<b>J</b> ratio appears to approach 1:1, reflecting the presence of the <b>K</b>α(1→3)<b>J</b>α(1→ disaccharide. Considerable overlap of the <b>K</b> and <b>J</b> anomeric signals appears to result in some inaccuracy in the integration, as the HSQC spectra demonstrate that both α- and β- reducing end anomeric protons were absent following borodeuteride reduction (see <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0183115#pone.0183115.s004" target="_blank">S4 Fig</a></b>).</p