The details of how the mucus hydrogel forms from its primary structural component, mucin polymers, remain incompletely resolved. To explore this, we use a combination of macrorheology and single particle tracking to investigate the bulk and microscopic mechanical properties of reconstituted MUC5AC mucin gels. We nd that analyses of thermal fluctuations on the length scale of the micron sized particles are not predictive of the linear viscoelastic response of the mucin gels, and that taken together, the results from both techniques help to provide complementary insight into the structure of the network. In particular, we show that macroscopic stiffening of MUC5AC gels can be brought about in different ways by targeting specic associations within the network using environmental triggers such as modications to the pH, surfactant, and salt concentration. Our work may be important for understanding how environmental factors, including pathogens and therapeutic agents, alter the mechanical properties of fully-constituted mucus.