In the NEQ approach, systematic errors can be introduced by starting NEQ transitions from local minima structures drawn from an incorrect or biased distribution (i.e. if the starting structures overpopulate some conformations and underpopulate others). In these cases, end state simulations can benefit from enhanced sampling techniques. In this work enhanced sampling methods were not investigated in detail in the NEQ approach, since we based this study on work of Gapsys et al. which has been successful without this. In hindsight, given our results (e.g. Figure ), it is worth exploring if faster convergence can be achieved with such enhancements. We tested enhanced end state sampling in the HSP90 system, but start-ing transitions from structures of the EQ HREX calculations did not improve results in this case.