The load partitioning between inter-phases and intragranular of austenite, their dislocation density evolution, and mechanical behavior were investigated in two medium Mn steels by in-situ high-energy synchrotron x-ray diffraction during tensile loading. The investigated steels were produced with different hot rolling parameters resulting in different initial microstructures. The initial microstructures were characterized using scanning electron microscope and electron backscatter diffraction analysis. Transmission electron microscope with high angle annular dark field technique was furthermore used to provide the fine microstructural features of deformed specimens in order to supplement the in-situ analysis. Despite the differences in the initial microstructure, the load partitioning from the softer ferrite to the stronger α’-martensite (α’) was observed; while different degrees of load partitioning for intragranular of γ occurred. The sample with a relatively low degree of load partitioning exhibits the intersection of two bundles of stacking faults (SF), one with high density SF, and the other with low density and limited SF, as well as the occurrence of α′ in the intersecting region. The secondary SFs and the relatively intermediate density of primary SFs bundle are characterized in γ for the sample with relatively high degree of load partitioning. The load partitioning between intragranular of γ is found to have a coordination relationship with the dislocation density evolution of γ; after the load partitioning, dislocation density of γ rapidly rises. The sample with intersection of two bundles SFs and the occurrence of α′, exhibits the higher transformation rate of γ into ε-martensite/α’, resulting in the larger work hardening rate. In addition, the early load on α’ is found to contribute to a higher yield strength. These results reveal the importance of assessing both phase-specifc strain/stress and the dislocation density evolution in order to predict the macroscopic mechanical properties of medium Mn steels.