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. 2016 Mar 17;35(8):866–880. doi: 10.15252/embj.201593596

Figure 2. USP19 interacts with Beclin‐1 complex member.

Figure 2

  • A
    293T cells were transfected with plasmids encoding HA‐USP19 and Flag‐tagged Beclin‐1 complex members (Flag‐ATG14L, Flag‐VPS15, Flag‐Beclin‐1, Flag‐VPS34, Flag‐UVRAG, and Flag‐AMBRA1), followed by immunoprecipitation (IP) with anti‐Flag beads and immunoblot analysis with anti‐HA. Throughout was the immunoblot analysis of whole‐cell lysates (WCL) without immunoprecipitation.
  • B, C
    Extracts of A549 cells (B) or human PBMCs (C) incubated with EBSS for various time points (above lanes) were subjected to immunoprecipitation with anti‐Beclin‐1 and immunoblot analysis with the indicated antibodies (shown on the left).
  • D, E
    HeLa cells were incubated in EBSS medium for 1 h after co‐transfection eGFP‐USP19 and dsRed‐Beclin‐1 for 48 h. Cells were imaged for eGFP and dsRed. Scale bar, 200 μm. Quantitative data are mean ± SEM of triplicate samples (20 cells per sample). ***P < 0.001 (two‐tailed Student's t‐test).
  • F
    Coimmunoprecipitation and immunoblot analysis of 293T cells transfected with deletion mutants of USP19 plasmid along with vector encoding Flag‐Beclin‐1.
  • G
    Coimmunoprecipitation and immunoblot analysis of 293T cells transfected with deletion mutants of Beclin‐1 plasmid along with vector encoding HA‐USP19.