Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 Jan 31;10:514. doi: 10.1038/s41467-019-08384-x

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

Fig. 6 — Enlarging the secretory lysosomes leads to enhanced NK cell function. a Confocal Z-stack showing MHC-I, LAMP-1, and granzyme B (GZMB) staining in primary NK cells following PIKfyve inhibition using overnight incubation with 1 μM vacuolin-1, 1 μM apilimod, or 1 μM YM201636. Scale bar is 5 μm. b Intracellular granzyme B expression in self-KIR⁺ and non-self-KIR⁺ NK cells following overnight incubation with the indicated PIKfyve inhibitor assessed by flow cytometry (n = 5 independent donors). c Representative example of a confocal image of primary NK cells treated overnight with 1 μM vacuolin-1 or DMSO. Scale bar is 2 μm. d Compiled confocal data on the volume of LAMP-1⁺ structures from cells treated overnight with DMSO or vacuolin-1 (n = 149 LAMP-1⁺ structures). e Cytosolic Ca²⁺-flux in NK cells in response to stimulation with biotinylated anti-DNAM-/2B4 (top) or anti-CD16 (bottom) crosslinked with streptavidin at the indicated timepoint. Cells were treated with 10 μM vacuolin-1 added directly before the assay and then maintained throughout the incubation time. f Representative FACS histogram of granzyme B versus CD107a expression following stimulation with K562 cells in the presence of DMSO or 10 μM vacuolin-1. g Frequency of CD107a^high+ and h IFNγ⁺ self-KIR⁺ and non-self-KIR⁺ NK cells after stimulation with K562 in the presence of DMSO or 10 μM vacuolin-1. i FACS-based killing assay showing NK cell killing of K562 cells after treatment with DMSO or 10 μM vacuolin-1 (n = 6 donors). j Relative phosphorylation of the indicated signaling molecules following stimulation with biotinylated anti-CD16 (10 μg/mL) crosslinked with streptavidin in the presence of 50 μM GPN or 10 μM vacuolin-1. Friedman’s test was used in panel (b). A non-paired t-test was used in panel (d). Paired t-test was used in panels (g–i). Whiskers show 5th to 95th percentile. Bars show the median. ****p < 0.0001; **p < 0.01; and *p < 0.05. Red and blue circles and box plots represent NK cells with self or non-self KIR, respectively. In panels (i) and (j), red and blue colors indicate cells treated with the indicated compounds