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. 2020 Dec 10;220(1):e202007052. doi: 10.1083/jcb.202007052

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© 2020 Zhang and Seemann

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Figure S3. — Acute degradation of GRASP55 and/or 65 does not affect Golgi stack reformation after BFA washout. (A) Cells were treated with doxycycline (dox) and then incubated with IAA and nocodazole for an additional 2 h. BFA was then added for 30 min to disassemble Golgi stacks. BFA was removed, and nocodazole (plus dox and IAA) was maintained to allow the reformation of individual Golgi stacks. The cells were fixed at the indicated time points and immunostained for Golgin-97 (red) and Golgin-84 (green) and labeled for DNA (blue). Scale bar, 10 µm. (B) Quantitation of the number of Golgin-97–marked Golgi elements per cell from A. n = 3 with an average of >10 cells per time point. Error bars represent mean ± SD. (C) Quantitation of stack formation from A as the percentage of Golgin-97–labeled elements that associate with Golgin-84. n = 3 with >10 cells per time point per condition per experiment. Error bars represent mean ± SD.