That should be like...
<asp:RegularExpressionValidator runat="server" ID="RegularExpressionFileUpload" ControlToValidate="FileUploadControl" ErrorMessage="Image Files Only (.jpg, .bmp, .png, .gif)" SetFocusOnError="true" ValidationExpression='<%#RegularExpressionVal %>' />
And the variable in code behind...
public string RegularExpressionVal="/^(([a-zA-Z]:)|(\\{2}\w+)\$?)(\\(\w[\w].*))+(.jpeg|.JPEG|.gif|.GIF|.png|.PNG|.JPG|.jpg|.bitmap|.BITMAP)$/";
But I don't understand this approach, because you can directly define this in aspx page.