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. 2021 Oct 22;11:20946. doi: 10.1038/s41598-021-00450-z

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Evaluation of TBC1D10C’s effect on phagocytosis of Burkholderia cenocepacia. The effect of TBC1D10C elimination and overexpression on phagocytosis of B. cenocepacia was determined by CFU (colony forming unit) quantification in the gentamicine protection assay and flow cytometry at a MOI of 50 (multiplicity of infection). (a) Quantification of CFU (phagocytosed living bacteria at 1.5 h postinfection) in infected BMDMs (mice, n = 7); (b) quantification of CFU in infected Raw 264.7 transfected cells (control vector and TBC1D10C, respectively); (c) quantification of CFU in infected BMDMs at 5 h postinfection (comparison of phagocytosed, in (a) bacteria vs. bacteria survival); (d), quantification of CFU in infected Raw 264.7 transfected cells at 5 h-postinfection (control vector and TBC1D10C, respectively); (e) quantification of CFU in infected Raw 264.7 transfected cells (control vector and Myo1G; unrelated protein to TBC1D10C, respectively); (f) left: data of quantified events expressed as a percentage of BMDMs infected with transformed B. cenocepacia (MH1K-pDsRed) by flow cytometry, right, representative histogram of 2 independent experiments for (f)(BMDM/( +)PE is positive control for infection: MH1K-pDsRed); (g) left: data for quantified events expressed as a percentage of Raw 264.7 transfected macrophages infected with transformed B. cenocepacia (MH1K-pDsRed) by flow cytometry, right, representative histogram of 2 independent experiments for (g) (Raw 264.7 cells/pDsRed is positive control for infection; cells highly infected). Data were analyzed by the Mann–Whitney test (P < 0.05), and graphs show SEM.