Before revision:  We used our methodology to revisit O'Shaughnessy et al.  tunable synthetic MAPK cascade, in which they claim to have found a new source of ultrasensitivity: ultrasensitivity generated de novo, which arises due to cascade structure itself. In this respect, we showed that the system's ultrasensitivity in its single-step cascade did not come from a cascading effect but from a hidden first-order ultrasensitivity process in one of the cascade's layer. Our analysis also highlighted the impact of the detailed functional form of a module's response curve on the overall system's ultrasensitivity in cascade architectures. Local sensitivity features of the involved transfer functions were found to be of the uttermost importance in this kind of setting and could be at the core of non-trivial phenomenology associated to ultrasensitive motifs .
After revision:  We used our methodology to revisit O'Shaughnessy et al. tunable synthetic MAPK cascade, and showed how our methodology might help modelers to better understand modeling alternatives .