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. Author manuscript; available in PMC: 2020 Jul 22.
Published in final edited form as: Curr Biol. 2019 Jul 11;29(14):2339–2350.e5. doi: 10.1016/j.cub.2019.06.032

Figure 5. EPAC2 is the major homolog regulating GDMT nucleation.

Figure 5.

A-D) Stills from a time-lapse (time, seconds) of MIN6 cells expressing EB3-mEmerald (+TIPs) and TGN-RFP (Golgi) in low glucose (A-B) and high glucose for 4-6 minutes (C-D). Vehicle control (A and C), EPAC activator (B) or HJC0197 (D). GDMTs (arrows) and centrosome (asterisk). Last panel is a maximum intensity projection over time (overlay). E-F) MIN6 cell immunostained for EPAC2 (cyan) and Golgi (GCC185, red). Low glucose (E) or high glucose 5 minutes (F). Insets (yellow dotted line) of the Golgi region (bottom panels). G) Nucleation events per minute for DMSO (grey), EPAC activator (red) and HJC0197 (blue) treated cells in low and high glucose. 5-95% boxplots, p < 0.001, one-way ANOVA and Tukey’s multiple comparison test; n = 36-40 cells. HI) MIN6 cell immunostained for EPAC2 (cyan) and MTs (red). DMSO (Vehicle, H) or nocodazole (I). A-D are maximum intensity projections of three confocal z-planes and E-F and H-I are maximum intensity projections of whole-cell confocal z-stacks. See also Videos S4 and S5.