Figure 4.

CryoSTAC of in situ Poly-GA aggregates with proteasomes recruitment in neurons.

(a) and (b) Correlative cryo-light and cryoFIB/SEM of rat cortical neurons cultured on EM grids and transduced with (GA)175-GFP. SEM (a) and FIB (b) images were aligned and superimposed with the GFP signal from the cryo-LM image. (c) Cryo-TEM low magnification image of the lamella superimposed with the GFP signal. (d) A tomographic slice recorded in the area with GFP signal (white square in (c)). Red arrowheads mark a dense network of poly-GA-GFP. (e) 3D rendering of an aggregate within a neuron transduced with (GA)175-GFP showing different macromolecules found either within or at the periphery of the aggregate. (f) Subtomogram classification of 26S proteasomes reveals enrichment of substrate processing conformations. GS, ground state; SPS, substrate-processing state [7^••].