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. 1997 Aug 25;138(4):927–938. doi: 10.1083/jcb.138.4.927

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Reconstitution of Rho-induced effects after extended permeabilization using a pig brain cytosolic extract. Cells were permeabilized according to protocol 2 (i.e., permeabilized for 6 min in the presence of digitonin before addition of stimulus; see Materials and Methods) in the presence of (a and b) 25 μg/ml V14Rho, (c and d) 2 mg/ml pig brain extract, (e and f) 25 μg/ml V14Rho plus 2 mg/ml pig brain extract. F-actin in the permeabilized cells was visualized using rhodamine-conjugated phalloidin (a, c, and e) and focal adhesions with anti-vinculin antiserum (b, d, and f). Bar, 30 μm.