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. 2003 Jan;23(2):482–492. doi: 10.1128/MCB.23.2.482-492.2003

FIG. 8.

FIG. 8.

Constitutive alkaline phosphatase expression and activity in cells expressing EWS/FLI-1. (A) C2C12 cells expressing EWS/FLI-1 (EF-1) or vector control (neo) were analyzed for alkaline phosphatase (alk phos) expression by semiquantitative RT-PCR (top) while in GM. As a control for alkaline phosphatase expression, C2C12 cells were treated with BMP-2 and analyzed at the indicated day (D) number. To control for equivalent sample processing, the expression of the ribosome-associated protein L7 mRNA is shown at the bottom. (B) C2C12 cells expressing EWS/FLI-1 or control (neo) (top panel) and the human ES tumor cell lines LD, TC-71, and SS (bottom panel) were analyzed for alkaline phosphatase activity by alkaline phosphatase enzyme-based histochemistry. (C) C2C12 cells stably expressing V12 ras (V12) or the parental vector (P) were cultured in GM or for 3 days in DM and analyzed for the expression of alkaline phosphatase, MyoD, or myogenin by semiquantitative RT-PCR. As a control for alkaline phosphatase expression, C2C12 cells treated with BMP-2 are shown at the right. For comparison, the constitutive expression of alkaline phosphatase by C2C12-EWS/FLI-1 cells is also shown at the right. The expression of L7 is included as a control for sample integrity.