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. 2019 Apr 12;14(5):882–892. doi: 10.1021/acschembio.8b01016

Figure 7.

Figure 7

HaloPROTAC-E induces degradation of endosomally localized proteins. A. SGK3-Halo cells were treated for up to 24 h with 300 nM HaloPROTAC-E alone, or in combination with VPS34IN1 at 1 μM. Cells were lysed, and remaining SGK3-Halo was analyzed by Immunoblot. B–F. HEK293 cells were transfected for 24 h for SGK3-Halo expression, and treated for 8h with 1 μM HaloPROTAC-E in the presence or absence of 1 μM VPS34-IN1 before fixing and staining for HaloTag7 and endosomal marker Rab5. SGK3-Halo was overexpressed to facilitate detection and quantitation. Anti-HaloTag7 antibody was detected with Alexa Fluor-594 coupled secondary, and Rab5 by Alexa Fluor-488 coupled secondary antibodies.