Supplemental

Materials and Methods Animal models of cerebral hypoperfusion and therapeutic induction of collateral growth For evalution of the effects of G-CSF on cerebral arteriogenesis, two different models of cerebral hypoperfusion were used, three vessel occlusion (3-VO) in rats [1,2] and left common carotid artery occlusion (CCAO) in mice (Fig. 1A). 3-VO includes bilateral vertebral artery occlusion followed by ligation of the left common carotid artery as described previously [1,3]. To show relevance of 3-VO in rats and CCAO in mice for the induction of permanent hypoperfusion, CCAO was performed in an additional rat group (n = 6) and compared with 3-VO. Cerebral blood flow was measured by laser Doppler flowmetry. Anaesthesia was initiated with 50 mg/kg ketamine/4 mg/kg xylazine i.p. in rats and 100 mg/kg ketamine/10 mg/kg xylazine i.p. in mice and maintained with 1-2% isoflurane in oxygen p.i.. In all groups Ropivacaine (5 mg/kg; Naropin®, AstraZeneca) was infiltrated into the wounds against pain.