Figure 6.
A role of Tex2–TMEM55B interaction in retrograde transport of LE/lys. (A) Representative images of live HeLa cell expressing Lamp1-mCh (gray) upon expression of empty vector, GFP-TMEM55B, TMEM55B siRNA, Tex2-KO, and TMEM55B siRNA and Tex2-KO with white dash lines indicating cell shape. (B) Western blots showing the efficiency of siRNA-mediated TMEM55B depletion in control or Tex2 KO HeLa cells. (C–E) Representative images of live HeLa cells expressing Lamp1-mCh (blue), Halo-TMEM55B (magenta), along with either GFP-Tex2 (C), GFP-Tex2-NT (D), or GFP-Tex2-∆1-276 (E) with white dash lines indicating cell shape. (F) Schematic diagram showing the definition of perinuclear regions. (G) Percentage of cells with lysosome clustering at perinuclear region in control (18 cells), TMEM55B overexpression (16 cells), TMEM55B siRNA (14 cells), Tex2-KO (15 cells), TMEM55B siRNA and Tex2-KO (18 cells), TMEM55B and Tex2 (16 cells), TMEM55B and Tex2-NT (1-514) (17 cells), or TMEM55B and Tex2-∆1-276 (16 cells) from more than three independent assays. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. (H) Representative images of a live COS7 cell expressing ER-GFP (green) and Lamp1-mCh (magenta) with time-lapse images of a boxed region shown on the right, from more than three independent assays. Yellow arrows denote one LE/lys undergoing retrograde transport with frequent contact with the ER (Video 1; time interval: 1.63 s). (I) Representative images of a live COS7 cell expressing GFP-Tex2 (green), Halo-TMEM55B (magenta), and Lamp1-mCh (blue) with time-lapse images of a boxed region shown on the right, from more than three independent assays. Yellow arrows denote one LE/lys undergoing retrograde transport along with the Tex2-labeled ER (Video 2; time interval: 2.53 s). Scale bar, 10 μm in the whole cell images and 2 μm in the insets in A, C–E, H, and I. Source data are available for this figure: SourceData F6.

A role of Tex2–TMEM55B interaction in retrograde transport of LE/lys. (A) Representative images of live HeLa cell expressing Lamp1-mCh (gray) upon expression of empty vector, GFP-TMEM55B, TMEM55B siRNA, Tex2-KO, and TMEM55B siRNA and Tex2-KO with white dash lines indicating cell shape. (B) Western blots showing the efficiency of siRNA-mediated TMEM55B depletion in control or Tex2 KO HeLa cells. (C–E) Representative images of live HeLa cells expressing Lamp1-mCh (blue), Halo-TMEM55B (magenta), along with either GFP-Tex2 (C), GFP-Tex2-NT (D), or GFP-Tex2-∆1-276 (E) with white dash lines indicating cell shape. (F) Schematic diagram showing the definition of perinuclear regions. (G) Percentage of cells with lysosome clustering at perinuclear region in control (18 cells), TMEM55B overexpression (16 cells), TMEM55B siRNA (14 cells), Tex2-KO (15 cells), TMEM55B siRNA and Tex2-KO (18 cells), TMEM55B and Tex2 (16 cells), TMEM55B and Tex2-NT (1-514) (17 cells), or TMEM55B and Tex2-∆1-276 (16 cells) from more than three independent assays. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. (H) Representative images of a live COS7 cell expressing ER-GFP (green) and Lamp1-mCh (magenta) with time-lapse images of a boxed region shown on the right, from more than three independent assays. Yellow arrows denote one LE/lys undergoing retrograde transport with frequent contact with the ER (Video 1; time interval: 1.63 s). (I) Representative images of a live COS7 cell expressing GFP-Tex2 (green), Halo-TMEM55B (magenta), and Lamp1-mCh (blue) with time-lapse images of a boxed region shown on the right, from more than three independent assays. Yellow arrows denote one LE/lys undergoing retrograde transport along with the Tex2-labeled ER (Video 2; time interval: 2.53 s). Scale bar, 10 μm in the whole cell images and 2 μm in the insets in A, C–E, H, and I. Source data are available for this figure: SourceData F6.

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