Figure S1.
Endosomal localization of GFP-tagged SNX27, VPS35, and VPS35(AAA3KE). (A and B) Co-localization of SNX27-GFP with 2×FYVE-mCh without or with VPS34-IN1 treatment (2 μM, 3 h) in Ctrl (A) and KO-91 (B) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of SNX27-GFP (green) and 2×FYVE-mCh (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (C and D) Co-localization of SNX27-GFP with mCh-CD63 without or with VPS34-IN1 treatment (2 μM, 3 h) in Ctrl (C) and KO-91 (D) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of SNX27-GFP (green) and mCh-CD63 (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (E and F) Co-localization of VPS35-GFP with 2×FYVE-mCh without or with VPS34-IN1 treatment (2 μM, 3 h) in Ctrl (E) and KO-91 (F) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of VPS35-GFP (green) and 2×FYVE-mCh (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (G and H) Co-localization of VPS35-GFP with mCh-CD63 without or with VPS34-IN1 treatment (2 μM, 3 h) in Ctrl (G) and KO-91 (H) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of VPS35-GFP (green) and mCh-CD63 (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (I and J) Co-localization of GFP-VPS35 and GFP-VPS35(AAA3KE) with 2×FYVE-mCh in Ctrl (I) and KO-91 (J) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of GFP-VPS35/GFP-VPS35(AAA3KE) (green) and 2×FYVE-mCh (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (K and L) Co-localization of GFP-VPS35 and GFP-VPS35(AAA3KE) with mCh-CD63 in Ctrl (K) and KO-91 (L) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of GFP-VPS35/GFP-VPS35(AAA3KE) (green) and mCh-CD63 (magenta) along the arrows across the endosomes in the zoom images are shown on the right.

Endosomal localization of GFP-tagged SNX27, VPS35, and VPS35(AAA3KE). (A and B) Co-localization of SNX27-GFP with 2×FYVE-mCh without or with VPS34-IN1 treatment (2 μM, 3 h) in Ctrl (A) and KO-91 (B) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of SNX27-GFP (green) and 2×FYVE-mCh (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (C and D) Co-localization of SNX27-GFP with mCh-CD63 without or with VPS34-IN1 treatment (2 μM, 3 h) in Ctrl (C) and KO-91 (D) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of SNX27-GFP (green) and mCh-CD63 (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (E and F) Co-localization of VPS35-GFP with 2×FYVE-mCh without or with VPS34-IN1 treatment (2 μM, 3 h) in Ctrl (E) and KO-91 (F) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of VPS35-GFP (green) and 2×FYVE-mCh (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (G and H) Co-localization of VPS35-GFP with mCh-CD63 without or with VPS34-IN1 treatment (2 μM, 3 h) in Ctrl (G) and KO-91 (H) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of VPS35-GFP (green) and mCh-CD63 (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (I and J) Co-localization of GFP-VPS35 and GFP-VPS35(AAA3KE) with 2×FYVE-mCh in Ctrl (I) and KO-91 (J) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of GFP-VPS35/GFP-VPS35(AAA3KE) (green) and 2×FYVE-mCh (magenta) along the arrows across the endosomes in the zoom images are shown on the right. (K and L) Co-localization of GFP-VPS35 and GFP-VPS35(AAA3KE) with mCh-CD63 in Ctrl (K) and KO-91 (L) cells. Zoom images show magnified frames in the merged images. Bars, 5 µm. Quantifications of linear pixel fluorescence of GFP-VPS35/GFP-VPS35(AAA3KE) (green) and mCh-CD63 (magenta) along the arrows across the endosomes in the zoom images are shown on the right.

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