The variation of the electronic structure normal to 1D defects in quasi-freestanding MoS 2 , grown by molecular beam epitaxy, is investigated through high resolution scanning tunneling spectroscopy at 5 K. Strong upwards bending of valence and conduction bands towards the line defects is found for the 4|4E mirror twin boundary and island edges, but not for the 4|4P mirror twin boundary. Quantized energy levels in the valence band are observed wherever upwards band bending takes place. Focusing on the 1 arXiv:2007.06313v1 [cond-mat.mes-hall] 13 Jul 2020 common 4|4E mirror twin boundary, density functional theory calculations give an estimate of its charging, which agrees well with electrostatic modeling. We show that the line charge can also be assessed from the filling of the boundary-localized electronic band, whereby we provide a measurement of the theoretically predicted quantized polarization charge at MoS 2 mirror twin boundaries. These calculations elucidate the origin of band bending and charging at these 1D defects in MoS 2. The 4|4E mirror twin boundary not only impairs charge transport of electrons and holes due to band bending, but holes are additionally subject to a potential barrier, which is inferred from the independence of the quantized energy landscape on either side of the boundary. Keywords band bending, scanning tunnelling spectroscopy, MoS 2 , polarization charge, mirror twin boundary Coupled to the rise of MoS 2 and other transition metal dichalcogenide (TMDC) semiconductors as prospective two-dimensional (2D) device materials came the need to investigate their one-dimensional (1D) defect structures, e.g. grain boundaries (GBs). Depending on their structure, GBs impair device performance to differing degrees when positioned in the channel of a single layer MoS 2 field effect transistor. 1-4 It is thus evident that control of the type and concentration of GBs is of importance for device fabrication. Besides satisfying scientific curiosity, it therefore pays to understand their effect on band structure and charge carrier transport. The lowest energy GBs are those hardest to avoid during growth, as the energy penalty associated with their introduction is marginal. In the three-dimensional (3D) world, these low energy GBs are 2D stacking faults or twin planes. For the case of SiC devices such defects cause increased leakage current, reduced blocking voltage, and the degradation of bipolar devices. 5,6 In the world of 2D materials, the analog to twin planes is 1D mirror twin boundaries (MTBs). These structural defects have some surprising effects on the band structure of monolayer MoS 2 , to be investigated in this manuscript.