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. 2018 Jul 10;9(7):773. doi: 10.1038/s41419-018-0806-4

Fig. 7. 26S depletion induces cytosolic condensation and nuclear distortion.

Fig. 7

a Experimental strategy of CRISPR/Cas9 editing to tag the endogenous 20S subunit PSMB6 with YFP at its C terminus (the N and C terminus of the YFP sequence are shown by capital letters). b The 26S and 20S proteasomal complex activity of the PSMB6-YFP proteasomes was analyzed by native gel electrophoresis in both naive and PSMB6-YFP cells as described. The 20S complex is shifted a bit higher with the addition of YFP. c The level of the YFP proteasome complexes was examined by immunoblotting with anti-YFP or by YFP fluorescence (Cy3 filter) (d). e Cellular morphology of PSMB6-YFP 293 cells upon 26S depletion. Endogenous proteasomes are visualized by YFP, and nuclei are stained by DAPI. HEK293 PSMB6-YFP cells were transfected with siRNA targeting PSMD1 or with control siRNA targeting luciferase. PSMD1 levels in siRNA-transfected PSMB6-YFP HEK293 cells were measured by qPCR (f)