The objective of this study is to investigate the effect of long-term static bending on the conductive characteristics of indium tin oxide (ITO) thin film in flexible optoelectronics. Two types of substrate are considered, namely ITO on polyethylene naphthalate (ITO/PEN) and ITO on polyethylene terephthalate (ITO/PET). Electrical properties of the ITO/PEN and ITO/PET sheets are measured in situ under static bending at various radii of curvature. Experimental results indicate that no significant change in electrical resistance of the ITO/PEN and ITO/PET sheets is found for compressive bending after 1000 h at a curvature radius of 10 mm or larger. However, the ITO/PEN and ITO/PET sheets are seriously damaged under a tensile bending of 10 mm radius and 5 mm radius, respectively. The given ITO/PET sheet exhibits a greater resistance to long-term mechanical bending than the ITO/PEN one, which is attributed to the effect of stiffness and thickness of substrate. As the given PET substrate has a lower stiffness and thickness than the PEN one, ITO thin film in the ITO/PET sheet has a smaller stress given a bending radius. Consequently, a smaller extent of change in the electrical conductance of ITO thin film is found in the ITO/PET sheet.