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. 2018 Jan 18;19(3):e44799. doi: 10.15252/embr.201744799

Figure 5. SCFFBXL 13 polyubiquitylates CEP192 isoform 3.

Figure 5

  1. Detection of polyubiquitylated Myc‐tagged CEP192 (aa 1–630) following immunoprecipitation from U2OS cells. Cells were co‐transfected with HA‐tagged ubiquitin and either a non‐targeting siRNA (siGL) or two siRNAs against FBXL13 (1) and (2). An empty vector (Vector) was used as a negative control.
  2. Relative FBXL13 mRNA levels in U2OS samples collected in (A) measured by qPCR, following depletion with either a non‐targeting siRNA (siGL) or two siRNAs against FBXL13 (1) and (2) (mean ± SD from three qPCR triplicates).
  3. Detection of polyubiquitylated Flag‐tagged CEP192 (aa 1–630) following immunoprecipitation from HEK293T cells under denaturing conditions. Cells were co‐transfected with HA‐tagged ubiquitin and either a scrambled or FBXL13 shRNA.
  4. Detection of SKP1 binding to immunoprecipitated Flag‐tagged FBXL13 wild type (WT) or a mutant lacking the F‐box (FBXL13 ∆F‐box) from HEK293T cells. An empty vector (Vector) was used as a negative control.
  5. Detection of polyubiquitylated Myc‐tagged CEP192 (aa 1–630) following immunoprecipitation from HEK293T cells. Cells were co‐transfected with HA‐tagged ubiquitin either in the presence of FBXL13 wild type (WT) or a mutant lacking the F‐box (∆F‐box). An empty vector (Vector) was used as a negative control.