Fig. 5: The ATG2C domain of VPS13C detects lysosome-membrane damage.
a, Domain organization of full-length VPS13C (top) as well as the domains of VPS13C used for the experiments of this figure (bottom). b, Fluorescence images of live RPE1 cells expressing full-length VPS13CHalo or VPS13C-Δ(ATG2C-PH)Halo under basal conditions (left). Magnified view of a region of a RPE1 cells showing co-localization of VPS13C-Δ(ATG2C-PH)Halo with Lamp1–GFP (right). c, Cartoon depicting the proposed association of VPS13C and its deletion constructs with Rab7 on the surface of lysosomes. d, Fluorescence image of live RPE1 cells expressing mCh-ATG2CVPS13C labelled with LysoView 640 and LipidSpot 488 (a lipid droplet marker; right). Magnified views of the boxed region are provided (right). e, Predicted structure of the ATG2C domain of VPS13C based on AlphaFold3 (left) and high-power views of the amphipathic helices (right). f, Fluorescence images of live RPE1 cells expressing mCh–ATG2CVPS13C before and after treatment with 1 mM LLOMe (left). Boxed regions are shown at higher magnification demonstrating co-localization of mCh-ATG2CVPS13C with Lamp1–GFP after LLOMe treatment (right). g, Putative model illustrating how binding of the ATG2CVPS13C domain to the bilayer may release an auto-inhibitory conformation of VPS13C to allow its binding to Rab7 on lysosomal surfaces. h, Fluorescence images of live RPE1 cells expressing mCh–ATG2CVPS13C and GFP–OSBP after treatment with 1 mM LLOMe (left) or 20 nM OSW1 (right). Grey dashed lines in the images represent cell outlines. Insets: magnified views of the boxed regions. f,h, Lipid droplets are labelled with asterisks. b,d,f,h, The experiments were repeated three times with similar results.
