White layer (WL) formation in metal cutting is generally found to have negative effects on the corrosion and fatigue life of machined components. Nowadays, the mechanism of the WL formation has not been understood very well, especially about the contribution of the thermal and mechanical loadings generated by the cutting process on WL formation. The relationship between subsurface plastic strain caused by mechanical loadings and the formation of WLs is of our concern. To address this issue, WL formation in hard turning of AISI 52100 under dry and cryogenic cooling conditions is investigated by subsurface plastic strain measurement using the micro-grid technique, observed by scanning electron microscope (SEM). Due to the considerable low temperature, WL is mainly generated by the mechanical effect rather than the thermal one, and this hypothesis is supported by physically based finite element method (FEM) simulations. From the investigations, we discover the existing plastic strain threshold, which governs the occurrence of WL in hard turning of AISI 52100 steel under cryogenic cooling conditions.