Movie 1.

Temporal dynamics of AGS3 BMC assembly under pH stress. Time-lapse video indicating the transition of AGS3 from a diffuse non-homogeneous cytosolic distribution to BMC. HeLa cells stably expressing AGS3-GFP were exchanged into HBSS 14185052 medium that lacked the ability to buffer pH under standard incubation conditions as described in “Materials and Methods”. Sequential imaging was performed at 20-second intervals over a span of 16 minutes to capture the progressive assembly of AGS3 into BMCs. During this time period the medium pH decreased from 7.4 to 6.2 as determined with pH test strips (pH range 5 to 10). The transition of AGS3 from a diffuse non-homogeneous cytosolic distribution to BMCs is documented in an in vivo time-lapse video generated by compiling 48 images (3 images/sec) by the Zeiss ZEN software, which is a digital imaging and analysis suite for microscopy.