Actin Cytoskeleton Inhibition Assay

CK666 (182515, Calbiochem), its inactive control CK689 (182517, Calbiochem) and Latrunculin A (L5163, Sigma) were dissolved in dimethylsulfoxide (DMSO). For treatment with inhibitors, 10 dechorionated embryos/larvae were added in 1 ml E3 without methylene blue to a well of a 12 well plate and 1 ml of 2x inhibitor (containing final DMSO concentration of 1%) or vehicle (1% DMSO - final concentration) as control in E3 without methylene blue was added to it and mixed gently with a pipette. CK666 and CK689 treatments (100 µM) were carried out for 1 h whereas Latrunculin treatment (2 µM) was performed for 30 min which is similar to a previous study^{34 (link)}.