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. 2016 Jun 17;8(8):851–862. doi: 10.15252/emmm.201506047

Figure 2. USP9X stabilizes XIAP to antagonize mitotic cell death independent of MCL1.

Figure 2

  1. Immunoblot analysis of WT, Xiap −/−, Mcl1 −/−, or cIap2 −/− mouse embryonic fibroblasts that were lentivirally infected with shRNA constructs directed against a non‐relevant mRNA (Ctrl) or against Usp9X mRNA and treated with taxol as specified.
  2. Two‐dimensional cell cycle analysis (BrdU/PI) of cells described in (A). Sub‐G1 and G2/M fractions of cells were quantified and averaged with two additional, independent experiments (n = 3, ± SD; WT: **P = 0.0016; ***P = 0.0004; Mcl1 −/−: ***P = 0.0003; **P = 0.0017; cIap2 −/−: **P = 0.0043, Student's t‐test). Black bars exemplify shRNA Ctrl and white bars shRNA Usp9X samples.
  3. Immunoblot analysis of HeLa cells transfected with a FLAG‐tagged USP9X expression construct or empty vector (EV) and treated with taxol for the indicated times.
  4. Immunoblot analysis of HeLa cells transfected with a FLAG‐tagged XIAP expression construct or empty vector and treated as in (C).
  5. Two‐dimensional cell cycle analysis (BrdU/PI) of cells described in (C) and (D). Apoptotic indices represent ratios of sub‐G1 to G1/S cells and are shown for analyses at the indicated time points (n = 3, ± SD). **P = 0.0020; ***P = 0.0004; Student's t‐test.
  6. Immunoblot analysis of HeLa cells that were transfected with siRNA oligonucleotides directed against a non‐relevant mRNA (Ctrl) or against Xiap mRNA and treated with taxol as specified.

Source data are available online for this figure.