Figure 7. Potentiation of PKC action increases cofilin dynamics during 5-HT responses. (A and B) Cofilin dynamics during myosin II reactivation in 5-HT backgrounds. Cells were pretreated with blebbistatin (30 µM, 15 min) followed by addition of 5-HT (10 µM, 20 min; Myosin II inhibition, A and B, left columns). Blebbistatin was washed out in the continued presence of 5-HT for 15 min (Myosin II reactivation, A and B, right columns). (A) Maximal overtime projections of cofilin (top), cofilin association (middle), and dissociation (bottom). (B) Quantification of cofilin association (top) and dissociation (bottom) in front and rear growth cone regions showing that cofilin dynamics are independent of myosin II activity in 5-HT backgrounds under normal conditions. Data are mean (a.u.) ± SEM. Number of growth cones evaluated: four, pooled from four independent experiments. (C and D) Cofilin dynamics with PKC potentiation. Cells were pretreated with blebbistatin (30 µM) and DAGKi (5 µM) for 15 min, and then 5-HT (10 µM) was added for 20 min (C and D, left columns). Blebbistatin was washed out in the continued presence of 5-HT and DAGKi for 15 min (C and D, right columns). (D) When PKC action is potentiated, myosin II reactivation enhances cofilin dynamics during 5-HT responses. Data are mean (a.u.) ± SEM. Number of growth cones evaluated: three, pooled from three independent experiments. *, P < 0.001 with two-tailed paired t test; #, P < 0.05 with two-tailed paired t test; Alexa Fluor 568–cofilin was injected into cells. Cofilin association and dissociation maps (A and C) generated as in Fig. 5. Scale bars, 5 µm. For B and D, regions were demarcated as in Fig. S2 A. See also Video 5.
Figure 7.

Potentiation of PKC action increases cofilin dynamics during 5-HT responses. (A and B) Cofilin dynamics during myosin II reactivation in 5-HT backgrounds. Cells were pretreated with blebbistatin (30 µM, 15 min) followed by addition of 5-HT (10 µM, 20 min; Myosin II inhibition, A and B, left columns). Blebbistatin was washed out in the continued presence of 5-HT for 15 min (Myosin II reactivation, A and B, right columns). (A) Maximal overtime projections of cofilin (top), cofilin association (middle), and dissociation (bottom). (B) Quantification of cofilin association (top) and dissociation (bottom) in front and rear growth cone regions showing that cofilin dynamics are independent of myosin II activity in 5-HT backgrounds under normal conditions. Data are mean (a.u.) ± SEM. Number of growth cones evaluated: four, pooled from four independent experiments. (C and D) Cofilin dynamics with PKC potentiation. Cells were pretreated with blebbistatin (30 µM) and DAGKi (5 µM) for 15 min, and then 5-HT (10 µM) was added for 20 min (C and D, left columns). Blebbistatin was washed out in the continued presence of 5-HT and DAGKi for 15 min (C and D, right columns). (D) When PKC action is potentiated, myosin II reactivation enhances cofilin dynamics during 5-HT responses. Data are mean (a.u.) ± SEM. Number of growth cones evaluated: three, pooled from three independent experiments. *, P < 0.001 with two-tailed paired t test; #, P < 0.05 with two-tailed paired t test; Alexa Fluor 568–cofilin was injected into cells. Cofilin association and dissociation maps (A and C) generated as in Fig. 5. Scale bars, 5 µm. For B and D, regions were demarcated as in Fig. S2 A. See also Video 5.

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