heavy-atom distances between all non-redundant residues separated by two or more residues served as the network input resulting in 666 nodes. MD trajectories were trained using VAMP neural networks with a time lag of 60 ns, training ratio of 0.9, and a batch size of 1000 that learned discretization in three metastable states by performing a hierarchical decomposition of the state space (Figure ). Starting structures were selected from the metastable states as per the DeepWEST protocol followed by the WE simulation of 90 µs with a resampling time, τ of 40 ps. Therefore, the total simulation time for the DeepWEST method was 120 µs. CVs for WE simulations were set to be mass-weighted root-mean-square deviation (RMSD) and mass-weighted radius of gyration (R g ) respectively. Both CVs were evenly spaced in bins of 0.05 nm ranging between [0 nm, 2 nm]. The target number of walkers per bin, n w was set to 8 for the WE simulation. Similarly, the conventional WE simulation was run for 130 µs with the same parameters (i.e. resampling time, CVs and binning parameters) as described previously for the DeepWEST simulation. The WE sim-ulations initiated from the unfolded state to the folded state with the CVs set to be the mass-weighted RMSD and mass-weighted R g respectively. Since the reference structure for computing RMSD is the initial unfolded state, RMSD is expected to increase during the simulation as the system folds. The unfolded region is defined as RMSD ≤ 0.60 nm, while the unfolded region is defined as RMSD ≥ 1.0 nm. For the same amount of simulation time, i.e., 130 µs, the DeepWEST method proved more effective than the conventional WE in covering the free energy landscape of a more complex protein, i.e., the NTL9 system. Figure and Figure represent the free energy landscapes for the NTL9 system for the DeepWEST and the conventional WE method, respectively, where the lowest energy state was set to zero. It is evident from Figure that the DeepWEST approach is more effective than the conventional WE in exploring other metastable states in the NTL9 system, particularly in the unfolded region. The rate constants between the unfolded and the folded states were not measured, but we expect the DeepWEST method to outperform the conventional WE method in kinetic rate measurements.