Figure EV3. ORF6 interacts with BAP31 and USE1.

• A
  GFP or GFP tagged ER membrane proteins were co‐expressed in HEK293T cells with ORF6‐Flag. Protein interactions were analyzed by immunoprecipitation with anti‐GFP beads and immunoblotting analysis.
• B
  Vector were co‐expressed with mCheey‐RAMP4. Cells were treated with 200 μM OA for 12 h, and then fixed. The ER was visualized with mCheey‐RAMP4 (red). LDs were labeled with LipidTOX Deep Red (white). Cells were imaged by confocal microscopy. Scale bar represents 10 μm.
• C
  Cells co‐expressing the ORF6‐Flag with mCheery tagged RAMP4 were transfected with BAP31 or USE1 siRNA for 36 h and treated with 200 μM OA for 12 h, then fixed and stained with anti‐Flag (green). LDs were labeled with LipidTOX Deep Red (blue). Cells were imaged by confocal microscopy. Scale bar represents 10 μm. Quantification of average number of LD‐ER contacts per LD. 12 cells (Negative) and 25 cells (BAP31/USE1 KD) from three independent experiments were calculated. Two‐tailed Unpaired Student's t‐test, ****P < 0.0001, ns means no significance. Error bars represent the mean ± SD.
• D
  Cell viability was analyzed in Fig 4L, three independent experiments. Two‐tailed Unpaired Student's t‐test, ns means no significance. Error bars represent the mean ± SD.

Source data are available online for this figure.