File:Uptake of MIF via CPZ sensitive endocytosis.png
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β-Arrestin1 Mediates the Endocytosis and Functions of Macrophage Migration Inhibitory Factor
FIGURE 1: Uptake of MIF via chlorpromazine (CPZ)-sensitive endocytosis. A, Uptake of MIF in RAW264.7 cells. RAW264.7 cells were incubated with MIF-TRITC for 1 h at 37°C, and then stained with LysoTraker Green and Hoechst. Subcellular distribution of MIF-TRITC was imaged with confocal microscopy. B, Uptake of MIF-TRITC is clathrin-dependent. RAWRAW264.7 cells were pretreated with Filipin (5 µM) or CPZ (5 µg/ml) for 30 min before stimulation with MIF-TRITC for another 2 h. Staining and confocal microscopy was carried out as in A. C, Uptake of MIF-TRITC is specific and temperature dependent. RAWRAW264.7 cells were treated with MIF-TRITC under the conditions indicated. MIF endocytosis was measured by FACS and the geometric mean of fluorescent signals was calculated by normalizing the signal of each sample with the signal of untreated cells. The experiment was repeated three times and means ± S.D. are indicated. *: Statistical analysis was carried out using the Student's t-test and P<0.05 was considered statistically significant. D, The sustained ERK activation induced by MIF is endocytosis-dependent. Serum-starved RAW264.7 cells were treated with CPZ for 30 min before stimulation with MIF for 15 min or 240 min. Western blot analysis was carried out using anti-ERK and anti-pERK antibodies. Scale bars: 10 µm. doi:10.1371/journal.pone.0016428.g001
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|current||13:10, 3 April 2014||3,894 × 2,960 (741 KB)||Z3399239||Uptake_of_MIF_via_CPZ_sensitive_endocytosis.png http://www.plosone.org/article/info:doi/10.1371/journal.pone.0016428 β-Arrestin1 Mediates the Endocytosis and Functions of Macrophage Migration Inhibitory Factor FIGURE 1: Uptake of MIF via chlorproma...|