No, functions One & Two will only be defined when their respective defun expressions are evaluated, and such defun expressions will only be evaluated when the function Main is evaluated. When loading an AutoLISP file, the contents are merely evaluated as if they were pasted to the command line or console - the interpreter does not 'scan' for nested function definitions - in fact, the interpreter is entirely indifferent to the expressions being evaluated: you could happily load an AutoLISP file containing only princ expressions without defining any functions whatsoever.