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. 2017 Apr 5;18(5):797–808. doi: 10.15252/embr.201643270

Figure 2. USP26 regulates SMAD2 activity and binds to SMAD7.

Figure 2

  1. 293T cells expressing knockdown vectors targeting USP26 or control vector were treated with TGF‐β overnight. Whole‐cell extracts were probed with the indicated antibodies.
  2. 293T cells expressing GFP‐USP26, GFP‐USP26 C/S, or control vector were treated overnight with TGF‐β. Whole‐cell extracts were probed with the indicated antibodies.
  3. 293T cells were transfected as indicated with GFP‐USP26 and Flag‐tagged SMAD1, SMAD2, SMAD3, SMAD4, SMAD6, and SMAD7. After 48 h, cells were lysed and immunoprecipitated with anti‐Flag affinity resin. Whole‐cell extracts were probed with the indicated antibodies.
  4. 293T cells were transfected as indicated with GFP‐USP26 and Flag‐tagged SMAD6 and SMAD7. After 48 h, cells were lysed and immunoprecipitated with anti‐GFP affinity resin. Whole‐cell extracts were probed with the indicated antibodies.
  5. 293T cells were transfected with GFP‐USP26 and cells were lysed and immunoprecipitated with anti‐GFP affinity resin. Whole‐cell extracts were probed with SMAD7.
  6. 293T cells were transfected with GFP‐USP26 and cells were lysed and immunoprecipitated with anti‐GFP affinity resin. Whole‐cell extracts were probed with SMAD6.
  7. 293T cells transfected with FL‐SMAD7, pRS‐USP26, control vector, and HA‐tagged ubiquitin. Following immunoprecipitation of SMAD7, lysates were resolved by SDS–PAGE and probed with the indicated antibodies.
  8. 293T cells transfected with FL‐SMAD7, pRS‐USP26, control vector, and wild‐type HA‐tagged ubiquitin or K48‐ubiquitin or K63‐ubiquitin mutants. Following immunoprecipitation of SMAD7, lysates were resolved by SDS–PAGE and probed with the indicated antibodies.

Source data are available online for this figure.