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. 2019 Jun 13;10:2586. doi: 10.1038/s41467-019-10556-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Environmental acidification controls speB expression. a Wild-type (WT) GAS was grown in THY broth, samples were collected at the indicated time points, and growth medium pH, speB transcript levels, and absorption at wavelength 600 nm (A₆₀₀) were determined. Right Y-axes represent fold-change in speB transcript levels (red) and A₆₀₀ (green). Fold-changes in transcript levels at indicated time points relative to starting culture (time point t = 0 h) are shown. Data are mean + standard deviation for three biological replicates. b WT GAS was grown in THY to late-exponential growth phase (LE, A₆₀₀ ~1.5), harvested by centrifugation, suspended in fresh THY adjusted to indicated pH and incubated for 1 h. The fold-change in speB transcript levels relative to WT-LE growth is shown. P values (*P < 0.5, ***P < 0.001) of the indicated samples relative to WT LE growth are shown. c Gross analyses of hindlimb lesions collected at 24 h postinfection from mice infected with 1 × 10⁷ CFUs of each indicated strain. Larger lesion with extensive tissue damage in WT-infected mice in pH 6 is boxed (black box). d Histopathology scores of mouse muscle tissue infected with each indicated strain (n = 3 per strain). Data are mean + standard deviation. P values (n.s. = not significant) of the indicated strains were compared to WT GAS in pH 8. e Twenty mice were infected intramuscularly and mean colony-forming units (CFUs) recovered from the infected muscle tissue are shown. n.s indicates no statistical significance (P > 0.05). Data graphed are mean ± standard deviation. f Analysis of the speB transcript level in the intramuscular lesions from mice infected with indicated strains. Samples were collected at 24 h postinoculation from the lesions (n = 4 per strain) and analyzed in triplicate by qRT-PCR. The speB transcript levels in WT-LE (A₆₀₀ ~1.5) was used as a reference and fold-changes in speB transcript levels relative to the reference are shown. P values (****P < 0.0001) of the indicated strains were compared to WT GAS in pH 6. P values were determined by t test