Figure 7. Effects of LIMK1 on... | Rockefeller University Press

Figure 7.

$Figure 7. Effects of LIMK1 on NRG-induced G-actin loss and cell extension. (A) Images of YFP-actin in MCF-7 cells expressing CFP-LIMK1(WT) or -LIMK1(D460A). Bar, 20 µm. (B) Effect of LIMK1 on the mobile fraction of Dp-actin. MCF-7 cells cotransfected with Dp-actin and mDsRed-LIMK1(WT) (red) or -LIMK1(D460A) (blue) were subjected to single FDAP analysis. Time curves of fluorescence decay were fitted using the diffusion coefficient of Dp-actin (13.7 µm2/s; thin lines). Data are means ± SEM of 15 cells (LIMK1(WT)) and 16 cells (LIMK1(D460A)) from at least three independent experiments. Right panel shows the mobile fractions. *, P < 0.05. (C) Effect of LIMK1 on NRG-induced G-actin loss. MCF-7 cells coexpressing Dp-actin and mDsRed-LIMK1(D460A) or -LIMK1(WT) were treated with NRG and analyzed by s-FDAP. FDAP values were normalized to the total intensity of Dp-actin in the same area as in Fig. 1 E. (D) Effect of LIMK1 on NRG-induced cell area extension. Cell area was measured using DIC imaging. (E) Quantification of the relative FDAP before (0–2 min) and after (4–6 min) NRG stimulation, calculated from data in C. *, P < 0.05. (F) Quantification of the increase in the relative cell area before (0–2 min) and after (10–12 min) NRG stimulation, calculated from data in D. Data are means ± SEM of 16 cells (LIMK1(D460)) and 15 cells (LIMK1(WT)) from at least three independent experiments. *, P < 0.05.$

Effects of LIMK1 on NRG-induced G-actin loss and cell extension. (A) Images of YFP-actin in MCF-7 cells expressing CFP-LIMK1(WT) or -LIMK1(D460A). Bar, 20 µm. (B) Effect of LIMK1 on the mobile fraction of Dp-actin. MCF-7 cells cotransfected with Dp-actin and mDsRed-LIMK1(WT) (red) or -LIMK1(D460A) (blue) were subjected to single FDAP analysis. Time curves of fluorescence decay were fitted using the diffusion coefficient of Dp-actin (13.7 µm²/s; thin lines). Data are means ± SEM of 15 cells (LIMK1(WT)) and 16 cells (LIMK1(D460A)) from at least three independent experiments. Right panel shows the mobile fractions. *, P < 0.05. (C) Effect of LIMK1 on NRG-induced G-actin loss. MCF-7 cells coexpressing Dp-actin and mDsRed-LIMK1(D460A) or -LIMK1(WT) were treated with NRG and analyzed by s-FDAP. FDAP values were normalized to the total intensity of Dp-actin in the same area as in Fig. 1 E. (D) Effect of LIMK1 on NRG-induced cell area extension. Cell area was measured using DIC imaging. (E) Quantification of the relative FDAP before (0–2 min) and after (4–6 min) NRG stimulation, calculated from data in C. *, P < 0.05. (F) Quantification of the increase in the relative cell area before (0–2 min) and after (10–12 min) NRG stimulation, calculated from data in D. Data are means ± SEM of 16 cells (LIMK1(D460)) and 15 cells (LIMK1(WT)) from at least three independent experiments. *, P < 0.05.