Interface charge transfer and electron-phonon coupling have been suggested to play a crucial role in the recently discovered high-temperature superconductivity of single unit-cell FeSe films on SrTiO3. However, their origin remains elusive. Here, using ultraviolet photoemission spectroscopy and element-sensitive X-ray photoemission spectroscopy, we identify the strengthened Ti-O bond that contributes to the interface enhanced electron-phonon coupling and unveil the band bending at the FeSe/SrTiO3 interface that leads to the charge transfer from SrTiO3 to FeSe films. We also observe band renormalization that accompanies the onset of superconductivity. Our results not only provide valuable insights into the mechanism of the interface-enhanced superconductivity, but also point out a promising route toward designing novel superconductors in heterostructures with band bending-induced charge transfer and interfacial enhanced electron-phonon coupling.The origin of interface charge transfer and electron-phonon coupling in single unit-cell FeSe on SrTiO3 remains elusive. Here, Zhang et al. report strengthened Ti-O bond and band bending at the FeSe/SrTiO3 interface, which leads to several important processes.