Figure 7.
Selective properties of SHIP164 vesicle clusters. (A) Structures of the chorein motif proteins SHIP164, its paralog UHRF1BP1, ATG2A, and VPS13C at the same scale as predicted by the fold prediction algorithm AlphaFold. (B) Live fluorescence image of an endogenously tagged SHIP164^mNG (green) HeLa cell expressing exogenous ATG9A-mScarlet (magenta). Scale bar, 5 µm. High magnification of the indicated regions is shown at right. Arrowheads indicate potential overlapping fluorescence. Scale bar, 1 µm. (C) Live fluorescence (inverted grays) image of a COS-7 cell expressing exogenous ATG9A-GFP (left) and SHIP164^mScarlet expression (right). Scale bar, 10 µm. High-magnification scale bar, 1 µm. (D) Fluorescence image of a fixed COS-7 cell expressing exogenous SHIP164^mScarlet (magenta) and immunolabeled with antibodies against ATG9A (green) and WIPI2 (shown in high magnification). Scale bar, 10 µm. High-magnification scale bar, 1 µm. (E) Live image of the cytoplasm of a COS-7 cell expressing exogenous SHIP164-GFP (magenta) and the autophagosome marker RFP-LC3 (green) in either complete medium (top) or starvation conditions (bottom). Note the lack of colocalization between these proteins. Scale bar, 4 µm. (F) Western blot (in kD) of RPE1 cells for SHIP164, LC3, and for tubulin as a loading control, either treated with Bafilomycin A1 or DMSO demonstrating normal autophagic flux in SHIP164 KO cells. Source data are available for this figure: SourceData F7.

Selective properties of SHIP164 vesicle clusters. (A) Structures of the chorein motif proteins SHIP164, its paralog UHRF1BP1, ATG2A, and VPS13C at the same scale as predicted by the fold prediction algorithm AlphaFold. (B) Live fluorescence image of an endogenously tagged SHIP164^mNG (green) HeLa cell expressing exogenous ATG9A-mScarlet (magenta). Scale bar, 5 µm. High magnification of the indicated regions is shown at right. Arrowheads indicate potential overlapping fluorescence. Scale bar, 1 µm. (C) Live fluorescence (inverted grays) image of a COS-7 cell expressing exogenous ATG9A-GFP (left) and SHIP164^mScarlet expression (right). Scale bar, 10 µm. High-magnification scale bar, 1 µm. (D) Fluorescence image of a fixed COS-7 cell expressing exogenous SHIP164^mScarlet (magenta) and immunolabeled with antibodies against ATG9A (green) and WIPI2 (shown in high magnification). Scale bar, 10 µm. High-magnification scale bar, 1 µm. (E) Live image of the cytoplasm of a COS-7 cell expressing exogenous SHIP164-GFP (magenta) and the autophagosome marker RFP-LC3 (green) in either complete medium (top) or starvation conditions (bottom). Note the lack of colocalization between these proteins. Scale bar, 4 µm. (F) Western blot (in kD) of RPE1 cells for SHIP164, LC3, and for tubulin as a loading control, either treated with Bafilomycin A1 or DMSO demonstrating normal autophagic flux in SHIP164 KO cells. Source data are available for this figure: SourceData F7.

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