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Inhibition of macropinocytosis prevents Alexa488-BSA uptake by rat pulmonary endothelial cells.

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posted on 2013-11-27, 02:43 authored by Hui-Hua Li, Jin Li, Karla J. Wasserloos, Callen Wallace, Mara G. Sullivan, Philip M. Bauer, Donna B. Stolz, Janet S. Lee, Simon C. Watkins, Claudette M. St Croix, Bruce R. Pitt, Li-Ming Zhang

(A) RPMEC were serum starved for 4 h followed by pretreatment with various inhibitors of macropinocytosis for 30 min including amiloride (Na/H exchange inhibitor, 3 mM), Cytochalasin D (actin polymerization inhibitor, 10 µM), Gö6976 (PKC inhibitor, 10 µM), then cells were incubated with Alexa488-BSA (50 µg/ml) for 15 min and fixed with 2% paraformaldehyde for imaging. Images were taken by confocal microscopy (40× magnification). Representative images from three independent experiments are shown. (B) The fluorescence intensity was determined by Imaris software with data are present as mean ± SEM from three independent experiments. Significant differences from control (** P < 0.01, *** P < 0.001) were determined by one-way ANOVA followed by Tukey's multiple comparisons.