LOXL2 is expressed by ECs in developmental and pathologic angiogenesis. (A) Immunofluorescence was performed on cryosections of P6 rat eye for detection of LOXL2 and either von Willebrand factor (VWF) or collagen IV (Col IV). Vitreous humor is indicated by asterisks and white lines. Bar: 20 μm. (B-D) Unilateral critical ischemia (I: ischemic) was analyzed 6 days postsurgery, using contralateral hindlimb as control (NI: non ischemic). (B) Fifty micrograms of total protein extracts were subjected to immunoblotting of LOXL2 (left panel). LOXL2 amount was quantified in extracts from 3 mice (right panel). Results are represented ± SD. (C) Quantification of loxl2 mRNA induction by real-time quantitative PCR. For each mouse (n = 3), actin was used as control. Mean relative induction in ischemic muscle (I) is represented ± SD. (D) In situ hybridization of loxl2 mRNA was performed on sections of non ischemic or ischemic tibialis anterior muscle (top lane). Slides were counterstained with toluidine blue. Serial sections were stained with H&E (middle lane), and CD31 was immunostained (bottom lane). Arrows indicate colocalization of loxl2 and CD31 in high magnification images (right panel). Bar: 200 μm. See also supplemental Figure 4.