In-situ electrochemical nanoindentation (ECNI) has been used to study the effect of hydrogen on the mechanical properties of austenitic stainless steel AISI 316L. Changing the electrode potential (via electrochemical charging) revealed the interconnected nature of the hydrogen effect on the nanomechanical properties of the stainless steel. At more positive cathodic potentials, a softening effect of hydrogen can be noticed, while significant hardening can be observed at more negative cathodic potentials. The hydrogen effects on the nanomechanical properties were analyzed in terms of the homogeneous dislocation nucleation (HDN) and the hydrogen-dislocation interactions from the energy point of view. The effects can be explained with the framework of the defactant theory and the hydrogen-enhanced localized plasticity (HELP) mechanism.