Xenbase Image ID: 82995

Fig. S3. Related to Fig. 3. Itpkb and InsP4 increase the levels of F-actin array after wounding and lead to an acceleration of the speed of wound healing in oocytes. (A�H) Oocytes were injected with 20 ng of mRNA encoding the F-actin probe, eGFP-DBmoesin, and coinjected with the respective mRNA or treated with the noted chemical. After 24 h, laser wounds were made and time-lapse confocal microscopy was taken. Time 0 s is equivalent to time after F-actin array formation. (A) F-actin array postwounding in control oocyte (F-actin probe alone) (Upper) or myc-itpkb oocyte (F-actin probe coinjected with 1ng myc-itpkb mRNA) (Lower). (Bars, 20 μm.) (B) F-actin array postwounding in oocyte incubated with 0.5 μM cIP4 (Upper) or 0.5 μM aIP4 (Lower). (Bars, 20 μm.) (C and D) Quantification of F-actin array intensity over time in the presence of itpkb (C) or aIP4 (D) relative to their respective controls. Each time point was analyzed by Student t test, nonparametric Mann-Whitney test. Results are shown as means � SEM. *P < 0.05. (E and F) Quantification of the relative F-actin ring diameter over time in the presence of Itpkb (E) or aIP4 (F) compared with their respective control. After testing Gaussian distribution by D�Agostino and Pearson omnibus normality test, statistic analysis was done by two-way ANOVA test. Results are shown as means � SEM. ***P < 0.001. (G) Speed of F-actin closure was calculated during the first 20 s, after the F-actin ring is first seen. (H) Quantification of speed rate in the presence of itpkb or aIP4, compared with their respective controls. Each time point was analyzed by Student t test, nonparametric Mann-Whitney test. Results are shown as means � SEM. *P < 0.05. Control n = 3, itpkb n = 5, cIP4 n=4,aIP4 n=4. Image published in: Soto X et al. (2013) Copyright © 2013. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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