We report on the electronic structure of freshly evaporated and air-exposed Molybdenum tri-oxide (MoO3) and the energy-level alignment between this compound and a hole-transport material [e.g., N,N′-diphenyl-N,N′-bis (1-naphthyl)-1,1′-biphenyl-4,4′-diamine (α-NPD)]. Ultraviolet and inverse photoelectron spectroscopy show that freshly evaporated MoO3 exhibits deep-lying electronic states with an electron affinity (EA) of 6.7 eV and ionization energy (IE) of 9.7 eV. Air exposure reduces EA and IE by ∼1 eV, to 5.5 and 8.6 eV, respectively, but does not affect the hole-injection efficiency, which is confirmed by device studies. Thus, MoO3 can be applied in low-vacuum environment, which is particularly important for low-cost manufacturing processes. Our findings of the energy-level alignment between MoO3 and α-NPD also leads to a revised interpretation of the charge-injection mechanism, whereby the hole-injection corresponds to an electron extraction from the organic highest-occupied molecular orbital (HOMO) level via the MoO3 conduction band.