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. 2019 Oct 8;10:4566. doi: 10.1038/s41467-019-12381-5

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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. 2 — KCs are critically involved in liver capture of disseminating C. neoformans. a Confocal microscopy revealed that KCs were completely depleted 24 h after treatment with clodronate liposomes (CLL). The control mice were treated with PBS liposomes (PBS lipo). KCs (labeled by Alexa Fluor 488 anti-F4/80 mAb): green; sinusoids (labeled by Alexa Fluor 647 anti-PECAM-1 mAb): red, pseudocolor. b Representative IVM images showing yeast cells (green, arrows) captured in the liver of mice treated with CLL or PBS lipo 30 min after i.v. injection with 100 × 10⁶ GFP-expressing C. neoformans H99. c C. neoformans (5 × 10⁶) was injected into the tail vein of mice (n = 4 mice/group) treated with CLL or PBS lipo. CFU in the liver was enumerated 10 min and 3 h post infection. d Vascular clearance and the half-life of circulating C. neoformans at various time points after injection of 100 × 10⁶ yeast cells into the tail vein of mice (n = 5 mice/group) treated with CLL or PBS lipo by analysis of IVM videos. e Blood CFU was enumerated in mice (n = 4 mice/group) treated with CLL or PBS lipo 10 min after i.v. infection of 5 × 10⁶ C. neoformans. f The possibility of circulating C. neoformans being captured was calculated by analysis of IVM videos from infected mice (n = 5 mice/group) treated with CLL or PBS lipo. g The possibility of trapped C. neoformans being released back to circulation was calculated by analysis of IVM videos from infected mice (n = 5 mice/group) treated with CLL or PBS lipo. h CFU in different organs was enumerated 3 h after i.v. injection of 5 × 10⁶ C. neoformans into the tail vein of mice (n = 4 mice/group) treated with CLL or PBS lipo. i CFU of different organs was enumerated 7 days after intratracheal infection with 1 × 10⁶ C. neoformans in mice (n = 5 mice/group) treated with CLL or PBS lipo. Data are expressed as mean ± SEM of three independent experiments. All data are from biologically distinct samples. Scale bars: 25 µm. *p < 0.05, **p < 0.01, ***p < 0.001, p values were calculated by two-way ANOVA (c, f, g) or Student’s t test (d, e, h, i). Source data are provided as a Source Data file