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. 1998 Sep;9(9):2561–2575. doi: 10.1091/mbc.9.9.2561

Figure 7.

Figure 7

Effect of transient expression of Rab GDI, Rab5DA, -5DN, -8DA, -8DN, -11DA, and -11DN on the TPA-induced disassembly and reassembly of stress fibers. wt MDCK cells were transfected with pEF-BOS-myc-Rab GDI, -L79Rab5 (Rab5DA), -N34Rab5 (Rab5DN), -L67Rab8 (Rab8DA), -N22Rab8 (Rab8DN), -L70Rab11 (Rab11DA), or -N25Rab11 (Rab11DN) using a lipofectAMINE reagent. At 24 h after the transfection, the cells were detached using an EDTA/trypsin solution, seeded onto 35-mm grid dishes, and further incubated in DMEM containing 10% FCS for 24 h. After the incubation, the cells were stimulated with 100 nM TPA and fixed at 15 min (A) or 2 h (B) after TPA stimulation. The cells were then double stained with rhodamine-phalloidin or the 9E10 anti-myc mAb to detect the transfected cells, and analyzed by confocal microscopy. (A) The percentage of the cells in which disassembly of stress fibers was observed at 15 min after TPA stimulation. (B) The percentage of the cells in which reassembly of stress fibers was observed at 2 h after TPA stimulation. None, untransfected cells; Rab GDI, the cells expressing myc-Rab GDI; Rab5DA, the cells expressing myc-L79Rab5; Rab5DN, the cells expressing myc-N34Rab5; Rab8DA, the cells expressing myc-L67Rab8; Rab8DN, the cells expressing myc-N22Rab8; Rab11DA, the cells expressing myc-L70Rab11; Rab11DN, the cells expressing myc-N25Rab11. The results shown are mean obtained from at least three independent experiments.