Figure 2.

Generation of Eph4 cell lines and characterization of the interferon-inducible system. (A) Construct used to generate the Eph4 stable lines overexpressing rab17 WT, rab17Q77L, and rab17N132I. The amino acid changes are indicated in shadow text. (B) Time course of protein expression after induction by interferon: protein expression from the Mx promoter was induced with 500 U of interferon. Protein extracts were prepared 0, 3.5, 10, 24, and 48 h after induction, blotted, and probed with a polyclonal antibody recognizing rab17. Finally, after induction for 24 h, interferon was washed out and cells were incubated for a further 24 h in the absence of interferon. (C) Concentration of interferon required for maximum protein expression: protein expression was induced with increasing units of interferon from 0 to 1,000 U per ml. Protein extracts were prepared 24 h after induction, blotted, and probed with a polyclonal antibody recognizing rab17.