Figure 3.
The N-terminal region of Tex2 is responsible for interacting with TMEM55B. (A) Left: representative images of COS7 cells expressing GFP-Tex2-∆TM (green) alone; middle: representative whole-cell images of a COS7 cell expressing GFP-Tex2-∆TM (green), Halo-TMEM55B (magenta), and Lamp1-mCh (blue); right: two enlarged images from the two boxed regions in the middle panel. (B–E) Representative images of COS7 cells expressing either GFP-Tex2-NT-1 (green; B), GFP-Tex2-NT-2 (green; C), GFP-Tex2 (277-517) (green; D), or GFP-Tex2-∆ (1-276) (green; E), along with Halo-TMEM55B (magenta) and Lamp1-mCh (blue) with insets. (F) Pearson’s correlation coefficient of TMEM55B vs. Tex2 mutants; Tex2-∆TM (10 cells); Tex2-1-540 (10 cells); Tex2-1-517 (10 cells); Tex2-277-517 (10 cells); and Tex2-∆1-276 (10 cells) in more than three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. (G) GFP-Trap assays demonstrate interactions between Halo-TMEM55B and Tex2 mutants in COS7 cells. (H) Pulldown assays using GFP-Tex2 bound on GFP-Trap beads and purified GST-TMEM55B demonstrate a direct interaction in a ceramide-independent manner. (I) As in H, pulldown assays using GFP-Tex2 bound on GFP-Trap beads and purified GST-TMEM55B demonstrate a direct interaction in a PS-independent manner. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in A–E. Source data are available for this figure: SourceData F3.

The N-terminal region of Tex2 is responsible for interacting with TMEM55B. (A) Left: representative images of COS7 cells expressing GFP-Tex2-∆TM (green) alone; middle: representative whole-cell images of a COS7 cell expressing GFP-Tex2-∆TM (green), Halo-TMEM55B (magenta), and Lamp1-mCh (blue); right: two enlarged images from the two boxed regions in the middle panel. (B–E) Representative images of COS7 cells expressing either GFP-Tex2-NT-1 (green; B), GFP-Tex2-NT-2 (green; C), GFP-Tex2 (277-517) (green; D), or GFP-Tex2-∆ (1-276) (green; E), along with Halo-TMEM55B (magenta) and Lamp1-mCh (blue) with insets. (F) Pearson’s correlation coefficient of TMEM55B vs. Tex2 mutants; Tex2-∆TM (10 cells); Tex2-1-540 (10 cells); Tex2-1-517 (10 cells); Tex2-277-517 (10 cells); and Tex2-∆1-276 (10 cells) in more than three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. (G) GFP-Trap assays demonstrate interactions between Halo-TMEM55B and Tex2 mutants in COS7 cells. (H) Pulldown assays using GFP-Tex2 bound on GFP-Trap beads and purified GST-TMEM55B demonstrate a direct interaction in a ceramide-independent manner. (I) As in H, pulldown assays using GFP-Tex2 bound on GFP-Trap beads and purified GST-TMEM55B demonstrate a direct interaction in a PS-independent manner. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in A–E. Source data are available for this figure: SourceData F3.

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