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. 2018 Apr 9;7:e32293. doi: 10.7554/eLife.32293

Figure 7. AIM-100 induced accumulation of YFP-HA-DAT in early but not recycling endosomes.

Figure 7.

(A) PAE/YFP-HA-DAT cells were transfected with CFP-Rab5, CFP-EEA.1, or mCherry (mCh)-Rab11, and incubated with 20 µM AIM-100 for 1.5 hr (CFP-Rab5 and CFP-EEA.1) or 2 hr (mCherry-Rab11) at 37°C. 3D images were acquired from fixed cells through 445 (CFP, red), 515 (YFP, green) and 561 nm (mCherry, red) channels. Individual confocal sections are presented for all except mCherry-Rab7 images where maximum intensity projection of z-stack of confocal images are presented to better demonstrate low extent of colocalization. (B) PAE/YFP-HA-DAT cells were incubated with 20 µM AIM-100 for 1.5 hr at 37°C, fixed and stained with the EEA.1 antibody followed by Cy3-conjugated secondary. 3D images were acquired through 515 (YFP, green) and 561 nm (Cy3, red) channels. Individual confocal sections are presented. Scale bars, 10 µm.