The influence of hot pressing on the pore structures of Nafion membranes was investigated by observing the Nafion before and after hot pressing with $^1H$ nuclear magnetic resonance (NMR) spectroscopy. The freezing point depression and chemical shift data of water in the Nafion indicated the presence of two different pore size ranges in Nafion. Hot pressing mainly reduced the sizes and number of the big pores. The reduction of water uptake and proton conductivity after hot pressing was explained by this variation of pore size and number. We demonstrated the potential application of chemical shift data and NMR cryoporometry experiments to measure the relative pore sizes, on a nano scale, and numbers.