Volume 121, Issue 5, Pages (November 2001)

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Date of download: 5/29/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Reduced Aquaporin 4 Expression in the Muscle Plasma.

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Volume 121, Issue 5, Pages 1203-1208 (November 2001) Expression and localization of the multidrug resistance proteins MRP2 and MRP3 in human gallbladder epithelia Daniel Rost, Jörg König, Gunter Weiss, Ernst Klar, Wolfgang Stremmel, Dietrich Keppler Gastroenterology Volume 121, Issue 5, Pages 1203-1208 (November 2001) DOI: 10.1053/gast.2001.28648 Copyright © 2001 American Gastroenterological Association Terms and Conditions

Fig. 1 Analysis of MRP1,MRP2, andMRP3 mRNA expression in human gallbladders (Gb). Reverse transcription was performed on deoxyribonuclease-digested total RNA with specific primers for MRP1, MRP2, and MRP3. For PCR analysis, primer pairs from human MRP1, MRP2, and MRP3 were used as described in Materials and Methods. Using the corresponding complementary DNA as the positive control, a 352–, 284–, and 452–base pair (bp) fragment, respectively, was obtained as expected. Amplification of a 661-bp fragment of β-actin was used as an internal control for the integrity of the isolated mRNA. A significant MRP2 and MRP3 mRNA expression was observed in all gallbladders. A weaker MRP1 mRNA expression was found in half of the gallbladder samples (Gb 2, 3, and 6). Mock = negative control omitting the template DNA. Gastroenterology 2001 121, 1203-1208DOI: (10.1053/gast.2001.28648) Copyright © 2001 American Gastroenterological Association Terms and Conditions

Fig. 2 Expression of (A and C) MRP2 and (D, F, G, and I) MRP3 in human gallbladder epithelial cells. Double-label immunofluorescence microscopy was performed as described in Materials and Methods. (A and C) Incubation of cryosections from human gallbladders with the MRP2-specific EAG5 antibody revealed MRP2 expression in epithelial cells. Apical localization was confirmed by (B and C) double staining for apical DPPIV resulting in (C) the yellow color of superimposed images. (D, F, G, and I) Incubation of cryosections with the FDS antibody raised against human MRP3 resulted in a basolateral staining of gallbladder epithelia. (E and F) MRP3 was absent in the apical membrane domain as shown by double immunofluorescence staining with apical DPPIV. Basolateral membrane domains were identified by double labeling gallbladder sections for MRP3 and desmoplakin (H and I; red fluorescence). Original magnification (A-F) 600×, (G-I) 1000×. Gastroenterology 2001 121, 1203-1208DOI: (10.1053/gast.2001.28648) Copyright © 2001 American Gastroenterological Association Terms and Conditions