Recently, research regarding non-pneumatic tires that are resistant to punctures has been actively conducted, and the spoke structure design of non-pneumatic tires has been found to be a crucial factor. This study aimed to analyze the vertical stiffness characteristics of a non-pneumatic tire based on the shape of the spoke under the application of a vertical load. The three-dimensional model of a commercial non-pneumatic tire was obtained from the manufacturer (Kumho Tire Co., Inc., Gwangju, Korea), and the vertical stiffness characteristics of the three tire models with modified spoke shapes were compared and analyzed based on a reference tire model. Results show that the vertical stiffness of the fillet applied model is most appropriate. Furthermore, the vertical stiffness characteristics of the analyzed tire models indicate that if fillets with a minimum size are applied to the spokes, the stability of the non-pneumatic tire is expected to improve.