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. 2000 Nov 6;192(9):1237–1248. doi: 10.1084/jem.192.9.1237

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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graphic file with name JEM000948.f5a.jpg — The effect of Nramp1 expression on Mn²⁺-induced quenching of internalized zymosan–FF6 fluorescence. Peritoneal macrophages from normal (+/+) and Nramp1 mutant (−/−) mice were plated for 48 h on coverslips, and allowed to ingest zymosan–FF6 particles for 30 min at 37°C in Ca²⁺-free medium containing 250 μM EGTA. Coverslips were mounted in thermoregulated chambers and examined using differential interference contrast optics to locate internalized particles. Fluorescence was measured with excitation at 360 nm before and after addition of 100 nM thapsigargin (thaps). Mn²⁺ (500 μM) was added to the cells where indicated and recording was resumed. A illustrates a typical experiment, and B summarizes the fractional quenching (as percentage of initial fluorescence) as a function of time. Data in B are means ± SE of 15 individual experiments for −/− macrophages (filled symbols) and 17 experiments for +/+ macrophages (open symbols).

graphic file with name JEM000948.f5b.jpg — The effect of Nramp1 expression on Mn²⁺-induced quenching of internalized zymosan–FF6 fluorescence. Peritoneal macrophages from normal (+/+) and Nramp1 mutant (−/−) mice were plated for 48 h on coverslips, and allowed to ingest zymosan–FF6 particles for 30 min at 37°C in Ca²⁺-free medium containing 250 μM EGTA. Coverslips were mounted in thermoregulated chambers and examined using differential interference contrast optics to locate internalized particles. Fluorescence was measured with excitation at 360 nm before and after addition of 100 nM thapsigargin (thaps). Mn²⁺ (500 μM) was added to the cells where indicated and recording was resumed. A illustrates a typical experiment, and B summarizes the fractional quenching (as percentage of initial fluorescence) as a function of time. Data in B are means ± SE of 15 individual experiments for −/− macrophages (filled symbols) and 17 experiments for +/+ macrophages (open symbols).