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

Figure 8.

Figure 8

Inhibition of the TPA-induced reassembly of stress fibers by transient expression of Rab5DN. wt MDCK cells were transfected with pEF-BOS-myc-L79Rab5 (Rab5DA) (a and b), -N34Rab5 (Rab5DN) (c and d), -L67Rab8 (Rab8DA) (e and f), -N22Rab8 (Rab8DN) (g and h), -L70Rab11 (Rab11DA) (i and j), and -N25Rab11 (Rab11DN) (k and l) 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 2 h after TPA stimulation. The cells were then double stained with rhodamine-phalloidin (a, c, e, g, i, and k) or the 9E10 anti-myc mAb (b, d, f, h, j, and l), and analyzed by confocal microscopy. Confocal images are shown at the basal levels. The results shown are representative of three independent experiments. Bar, 10 μm. Panels k and l show the Rab11DN-expressing cell in which the TPA-induced reassembly of stress fibers is observed, whereas the expression of Rab11DN inhibits their TPA-induced reassembly to a small extent (see Figure 7).