Figure 5. Functional analysis of in vitro hNMJ culture at day 20 of NMJ differentiation.

(A) Ca²⁺ transients and movements of the myotube were measured simultaneously in 2 regions of interest (ROIs). (B) Ca²⁺ imaging in 5 ROIs. Ca²⁺ transients increased with the application of calcium and stopped with the application of curare. (C) Motion analysis in 4 ROIs. (D) Motion analysis of myotubes before and after treatment with a gap junction inhibitor, GAP27. GAP27 was not washed out in the experiment. See also Supplemental Figure 9 and Supplemental Videos 1 and 2.