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. 2017 Jun 13;7:3431. doi: 10.1038/s41598-017-03477-3

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© The Author(s) 2017

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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OMS-mediated autophagy activation is essentially required for antimicrobial responses against mycobacterial infection in vitro and in vivo. (a) Atg7 ^fl/fl LysM-Cre⁻ and Atg7 ^fl/fl LysM-Cre⁺ BMDMs were infected with Mtb (moi = 10) for 4 h and then treated with OMS-A (10 μM) and OMS-B (10 μM). (b) Human MDMs were transduced with non-specific shRNA (shNS) or ATG5-specific shRNA (shATG5)-expressing lentivirus for 48 h and then infected with Mtb and then treated with OMS-A and OMS-B. (inset) RT-PCR analysis of ATG5 mRNA expression of transduction efficiency. (a and b) CFU assay. The intracellular bacterial loads were determined at 3 days after infection. (c and d) W¹¹¹⁸ flies were infected with M. marium (CFU = 500), followed by incubation with or without OMS-A (10 μM) in the presence or absence of chloroquine (1 μM). Positive control group was treated with AMK (1 μg/ml). (e) Live flies were counted at 24 h intervals (n = 40). The error bars indicate 95% confidence intervals. Log-rank analysis of the survival curves indicated that the susceptibility to M. marinum was significantly increased in the group of OMS-A with chloroquine, when compared with that of OMS-A only (***p < 0.001). (f) Each group (n = 20) of flies was harvested at 7 days, homogenized, and quantified by CFU assay. (e and f) Atg7 control and Atg7 mutant flies were infected with M. marium (CFU = 500), followed by incubation with or without OMS-A (10 μM). (e) Flies were counted at 24 h intervals (n = 50). The error bars indicate 95% confidence intervals. Log-rank analysis of the survival curves showed that the survival rates of Atg7 control flies were significantly increased by OMS-A treatment (***p < 0.001), whereas these were not significant in Atg7 mutant flies with the same treatment. (f) Each group (n = 20) of flies was harvested at 3 days, homogenized, and quantified by CFU assay. All data represent the means ± SD of triplicates from each sample. **p < 0.01, ***p < 0.001, compared with control conditions (a,b,d,f). SC, solvent control; AMK, amikacin; CQ, chloroquine.