English: Supplementary Movie 3 Time-lapse of the NMDAR-dependent downregulation of Venus-NRG2 clusters in response to glutamate. Neurons were infected at DIV3 with Venus-NRG2 and live-imaged on DIV 21 (see Methods). The movie depicts representative neurons shown in Figure 3, with individual frames acquired every 30 seconds. Baseline fluorescence was acquired for 5 min, either in the absence (S2) or presence (S3) of 50 μM AP5, followed by bath application of 20 μM glutamate for 10 min. In movie S2, Venus-NRG2 clusters dim immediately after the addition of glutamate and quickly fade to background levels. In movie S3, Venus-NRG2 fluorescence remains stable throughout the duration of the recording session due to the blockade of NMDA receptors by AP5.