Fig. 2: LC3b and poly-ubiquitin interactions with p62 filaments visualized by single-particle fluorescence microscopy and negative staining EM. | Nature Communications

Fig. 2: LC3b and poly-ubiquitin interactions with p62 filaments visualized by single-particle fluorescence microscopy and negative staining EM.

From: Structural organization of p62 filaments and the cellular ultrastructure of calcium-rich p62-enwrapped lipid droplet cargo

Fig. 2

p62 filament (a) labeled with Cy5 on a TIRF microscope and (b) in the presence of CFP-LC3b. c p62 condensates forming in the presence of GST-4xUbiquitin (GST-4xUb). d p62 condensates formed by GST-4xUb disintegrate after addition of LC3b. e Negative staining of p62 filaments. Dashed boxes correspond to an inset at higher magnification. f Negative staining of p62 filaments decorated with LC3b (sample as in B) (white arrows in inset). g Histograms of p62 filament length before and after addition of LC3b. h Negative staining of p62 condensates (sample as in D with GST-4xUb). i Negative staining of p62 condensates after addition of LC3b (sample as in D with GST-4xUb). j Illustration of revealed p62 domain interactions: The ZZ domain may potentially bind divalent cations Zn2+ and Ca2+, while interaction of LC3b via the LIR motif competes with the binding of ubiquitin to the UBA domain. All experiments were conducted three times (N = 3). Source data are provided as a Source Data file.

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