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. 2019 Mar 25;9:5052. doi: 10.1038/s41598-019-41513-6

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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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Increased Virulence of 10 Gen Old C. auris Cells from FLC- Susceptible S1: (a) Cell size and Cell wall thickness of the old (10 Gen) cells were significantly greater compared to young (0–3 Gen) cells. Cell wall thickness (n = 23) and Cell Size (n = 150) were measured by ImageJ software from the microscopic images of individual young and old cells. Student’s t-test was performed to analyze the significance (*p < 0.001). Transmission Electron Microscopic (TEM) image of the old (10 Gen) cells of C. auris showing increased cell wall thickness compared to the young (0–3 Gen) cells. (b) Percent Neutrophil-mediated Killing of young (0–3 Gen: black bar) and old (10 Gen: red bar) cells of C. auris. The assay was done in triplicate and error bars signify standard deviation. Student’s t-test with Welch’s correction was performed to determine the significance; *p = 0.0266. (c) Percent adhesion of young (0–3 Gen: black bar) and old (10 Gen: red bar) cells of C. auris. The assay was done in triplicate and error bars signify standard deviation. Student’s t-test with Welch’s correction was performed to determine the significance; *p = 0.0291. (d) qPCR analysis to study the expression of ALS5 in young and old cells. ACT1 gene was used as an internal control and the data was normalized to gene expression in young cells. The assay was done in triplicate and error bars signify standard deviation. Multiple t-test was performed using Holm-Sidak method to analyze the significance; *p = 0.016. (e) Virulence of young (0–3 Gen) and old (10 Gen) cells of C. auris in Galleria. 20 larvae were infected separately with 10⁵ young and old cells. Log rank test was used to determine the statistical significance.