Water evaporation-driven energy harvesting is an emerging mechanism for contributing to green energy production with low cost. Herein, we developed polyacrylonitrile (PAN) nanofiber-based evaporation-driven electricity generators (PEEGs) to confirm the feasibility of utilizing electrospun PAN nanofiber mats in an evaporation-driven energy harvesting system. However, PAN nanofiber mats require a support substrate to enhance its durability and stability when it is applied to an evaporation-driven energy generator, which could have additional effects on generation performance. Accordingly, various support substrates, including fiberglass, copper, stainless mesh, and fabric screen, were applied to PEEGs and examined to understand their potential impacts on electrical generation outputs. As a result, the PAN nanofiber mats were successfully converted to a hydrophilic material for an evaporation-driven generator by dip-coating them in nanocarbon black (NCB) solution. Furthermore, specific electrokinetic performance trends were investigated and the peak electricity outputs of Voc were recorded to be 150.8, 6.5, 2.4, and 215.9 mV, and Isc outputs were recorded to be 143.8, 60.5, 103.8, and 121.4 μA, from PEEGs with fiberglass, copper, stainless mesh, and fabric screen substrates, respectively. Therefore, the implications of this study would provide further perspectives on the developing evaporation-induced electricity devices based on nanofiber materials.