Movie S5

The effect of 2-APB (an IP3 receptor blocker) on neural rosette maintenance.
Neural rosettes were derived from human embryonic stem cells stably transfected with the pEF1a-AcGFP1-N1 vector. Time lapse imaging of a neural rosette responding to 50 μM 2-Aminoethoxydiphenyl borate (2-APB) was filmed before treatment for 10 min in N2B27 medium, under 2-APB treatment for 3 hr, and in N2B27 medium after washing off 2-APB overnight. The movie was extracted from a full-length film, which was recorded at 1 picture/5 min using the confocal microscope system (LSM 700, Carl Zeiss GmbH, Germany). The movie depicts the partial dismantlement of a neural rosette under 2-APB treatment for 3 hr. However, the destructed neural rosette failed to form new neural rosettes in N2B27 medium overnight after lifting 2-APB treatment.