Fig. 5. Overexpression of the mDia1 FH2 domain interferes with Golgi polarization in wound-edge NIH 3T3 cells. Golgi polarity, at the edge of the wound, of cells expressing full-length mDia1, mDia-deltaN3, mDia F2 and mDia-deltaN3KA3: Golgi reorientation to face the wound was evaluated when wound-edge NIH 3T3 cells were microinjected with the four different pEGFP-mDia1 expression constructs (see Fig. 4). (A) The microinjected cells expressing the GFP-fusion protein are shown (top panels): mDia-deltaN3 (left), mDia F2 (middle) and mDia-deltaN3KA3 (right). Cells were fixed and stained for the Golgi apparatus with the anti-ß-COP antibody (bottom panels). (B) The percentage of Golgi orientated towards the wound was determined at 3 hours post wounding for cells microinjected with the four mDia1 expression vectors. Golgi polarization, over the time course of wound closure, of non-microinjected wound-edge cells was included as a control (blue-grey bars) (see also legend on the right side of the graph). (C) Quantification data. The first number corresponds to the number of Golgi observed and the second, in parentheses, is the number of separate experiments. Bar, 40 µm.

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