Corresponding to Fig. 3. FLI-06 does not disrupt existing VSVG-EYFP-containing ERES. Hela cells transfected with VSVG-EYFP were incubated at 40°C over night, then treated with BFA (B) and nocodazole (N) for 20-30 min on ice. Cells were shifted to 32°C and incubated for 45 min to induce gERES. After 45 min incubation at 32°C, cells were incubated with BN or BN with 10 mM FLI-06 (BN+F). Images were taken every 5 sec on a Zeiss spinning disk confocal microscope. The arrow in the BN treated cell indicates an ERES with the typical increase-collapse-increase intensity profile, the arrowhead in the BN+F condition a collapsing ERES with no new cargo recruitment.