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Fig. 5. `Eat-each-other' endocytosis of claudins. (A,B) Time-lapse observation. Elapsed time is indicated at the bottom (in minutes:seconds). Eph4:NGFP-Cld3 cells were co-cultured with parental Eph4 cells, and a border between these two distinct types of cells was observed. In the first frame (A), four Eph4:NGFP-Cld3 cells (G) and four Eph4 cells (asterisks) were identified by fluorescence microscopy (left panel) and phase contrast image microscopy (right panel). From frame 35:00 to 70:30, one GFP-positive granule (arrowhead) budded off from the GFP-positive TJ, and moved into parental Eph4 cells. See Movie 3 (http://jcs.biologists.org/supplemental/). (C,D) Co-culture of MDCK transfectants exogenously expressing nontagged claudin-3 (MDCK-Cld3) or N-terminally FLAG-tagged claudin-1 (MDCK-FCld1). Parental MDCK cells expressed no endogenous claudin-3. Confluent cell sheets were double stained with anti-claudin-3 pAb and anti-FLAG mAb in green and red, respectively. As schematically depicted in C, the border between MDCK-Cld3 (Cld3) and MDCK-FCld1 (FLAG-Cld1) cells was focused on. Claudin-3-posive granules (arrowheads; green) were detected in MDCK-FCld1 cells, and FLAG-positive granules (arrows; red) were scattered in the cytoplasm of MDCK-Cld3 cells. Bars, 10 µm (A); 10 µm (B); 10 µm (D).
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