Accurate assessment of the mechanical properties of organic matter, clay matrix, and bulk shale during maturation remains a challenge. Here, we aim to assess the mechanical properties of organic-rich shale during maturation using a combination of nanoindentation methods and various geochemical analyses, i.e., mineral composition, mass loss rate, chemical structure of organic matter, and Rock-Eval analyses. Results show that the evolution of mechanical properties of organic matter in shale during maturation can be divided into: the main oil-generation stage, and the condensate oil and gas generation stage. The stiffening of organic matter in the shale is mainly due to increased aromaticity and condensation of aromatic groups. The clay matrix experiences a slight decrease in hardness and Young’s modulus at low maturity levels due to the generation of liquid hydrocarbons. However, overall, the clay matrix becomes stiffer as the shale matures due to shale dehydration, expulsion or cracking of liquid hydrocarbons, transformation of clay minerals, and hardening of organic matter. The Young’s modulus and hardness of bulk shale generally increase with increasing maturity. This is closely related to the hardening of organic matter and clay matrix, as well as the development of the more compact and dense microstructure in the shale.