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. 2010 Aug 2;30(19):4604–4615. doi: 10.1128/MCB.00197-10

FIG. 1.

FIG. 1.

mDia1 deficiency impairs v-Src-induced morphological transformation. (A) Generation of WT-NY72 and KO-NY72 cell populations. MEFs from WT and mDia1−/− (KO) littermates were immortalized with SV40 T antigen and then infected with retrovirus harboring ts-v-Src, NY72. Populations of cells stably expressing SV40 T antigen and NY72 were isolated by two-step drug selection. (B) Expression and activation of v-Src in WT-SV40, KO-SV40, WT-NY72, and KO-NY72 cells. The cells were maintained at 37°C. Cell lysates were prepared and probed for α-tubulin, mDia1, Src, and pY416-Src. Ratios of the densities of pSrc to Src bands are shown below the image. (C) Proliferation of WT-SV40, KO-SV40, WT-NY72, and KO-NY72 cells. Cells were seeded and cultured at 37°C in a 35-mm dish in DMEM containing 10% FBS and harvested at indicated times for cell counting. Means ± SEM of data from three independent experiments is shown. (D) v-Src-induced morphological changes in WT-NY72 and KO-NY72 cells. WT-NY72 and KO-NY72 cells were grown at either 40°C or 37°C and observed under phase-contrast microscopy. (E) Actin cytoskeleton in WT-NY72 and KO-NY72 cells. WT-NY72 and KO-NY72 cells were grown at either 40°C or 37°C, fixed, and stained with Oregon green-phalloidin. Bars in panels D and E, 50 μm.