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[Preprint]. 2023 Aug 28:2023.08.28.555157. [Version 1] doi: 10.1101/2023.08.28.555157

Figure 2: CHMP4B, but not Galectin-3 is recruited to fibril-containing lysosomes.

Figure 2:

TMEM192-RFP expressing astrocytes were treated with labeled PFF to verify endosomal uptake (a). These same cells were additionally transduced with CHMP4B-HALO to monitor potential ESCRT recruitment to fibril-containing lysosomes. At rest, CHMP4B was mostly diffuse with few puncta that rarely interacted with TMEM192-positive lysosomes (b). Treatment with labeled PFF triggered the robust formation of CHMP4B puncta that overlapped with fibrils and lysosomes (c-d). (e) Quantification of CHMP4B-expressing cells before and after PFF treatment (n=3 replicates, 20 random and independent fields of 10–20 cells per replicate, n=386 cells, represented as mean and standard deviation across fields). TMEM192-RFP expressing astrocytes were transduced with Gal3-GFP and the fractional overlap of GFP puncta area and endogenous TMEM192-RFP area was compared before and after fibril treatment (f-h) (n=4, 20 random and independent fields of 10–20 cells per replicate, n=372 cells). Scale bars: 10 µm.