Background and purpose We compared two imaging biomarkers for dose-escalation in patients with advanced non-small cell lung cancer (NSCLC). Treatment plans boosting metabolically active sub-volumes defined by FDG-PET or hypoxic sub-volumes defined by HX4-PET were compared with boosting the entire tumour. Materials and methods Ten NSCLC patients underwent FDG- and HX4-PET/CT scans prior to radiotherapy. Three isotoxic dose-escalation plans were compared per patient: plan A, boosting the primary tumour (PTV_prim); plan B, boosting sub-volume with FDG >50% SUV_max (PTV_FDG); plan C, boosting hypoxic volume with HX4 tumour-to-background >1.4 (PTV_HX4). Results Average boost volumes were 507 ± 466 cm³ for PTV_prim, 173 ± 127 cm³ for PTV_FDG and 114 ± 73 cm³ for PTV_HX4. The smaller PTV_HX4 overlapped on average 87 ± 16% with PTV_FDG. Prescribed dose was escalated to 87 ± 10 Gy for PTV_prim, 107 ± 20 Gy for PTV_FDG, and 117 ± 15 Gy for PTV_HX4, with comparable doses to the relevant organs-at-risk (OAR). Treatment plans are available online (https://www.cancerdata.org/10.1016/j.radonc.2015.07.013). Conclusions Dose escalation based on metabolic sub-volumes, hypoxic sub-volumes and the entire tumour is feasible. Highest dose was achieved for hypoxia plans, without increasing dose to OAR. For most patients, boosting the metabolic sub-volume also resulted in boosting the hypoxic volume, although to a lower dose, but not vice versa.