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. 2000 Dec 19;97(26):14394–14399. doi: 10.1073/pnas.97.26.14394

Figure 3.

Figure 3

Ski represses BMP signaling in mammalian cells. (A) Ski inhibits BMP-induced ALP in W-20-17 cells. W-20-17 cells plated in a 96-well cluster were transfected with increasing amounts of c-ski as indicated. Twenty-four hours after transfection, the cells were treated with 250 ng/ml BMP2 for 40 h. ALP was measured by an Elisa reader and expressed as p-nitrophenol produced in nmol/min per mg protein. Ski represses BMP-induced transcriptional activation in W-20-17 (B) and Hep3B (C and D) cells. W-20-17 and Hep3B cells were cotransfected with 0.75 μg of 15xGCCG-Luc (B and C) or Xvent2-Luc (D), 0.5 μg of constitutive active type I receptors (Alk3* or Alk5*), 0.75 μg of ski, and 0.15 μg of Smads. Luciferase activity was measured 48 h later. Alk3*, (Q233D)Alk3; Alk5*, (T204D)Alk5; S1, Smad1; S4, Smad4.