García-Quintans et al.
SupplementaRY Information
Results
In order to evaluate the impact of PGC-1 loss in the number of mitochondria in vascular endothelial cells, we analized mitochondrial number in wild-type and PGC-1-/- MLEC labeling the mitochondria with an antibody directed against the outer-membrane mitochondrial protein TOMM22. We found that, the number of mitochondria was significantly reduced in PGC-1-/- MLEC compared to wild-type cells. Furthermore, the mitochondria of PGC-1-/- MLEC showed a more marked perinuclear distribution, a characteristic generally associated with low oxidative capacity and high ROS generation rates [1].
Figure Captions
Supp. Fig. 1.-The graph shows the average lacurarity of the inner vascular plexus of retinas from control and OIR mice. Data are means ±SD (*) p ≤ 0.05 vs. Control.
Supp. Fig. 2.-qPCR determination of mRNA leves of the indicated genes from total retinal preparations from wild-type and PGC-1-/- mice. Data are means ±SD (*) p ≤ 0.05 vs. Control.
Supp. Fig. 3.-The graphs show the average cross-section area, perimeter and maior axis of vessels from matrigel subcutaneous implants. Data are means ±SD (*) p ≤ 0.05 vs. Control.
Supp. Fig. 4.-Flat mount retinas from PGC-1-/- mice immunostained with IsoB4 show reduded stain in OIR mice.
Supp. Fig. 5.-Confocal microscope images of OIR flat mount retinas immunostained with VE-cadherin (60Xobjective) show diffuse stain in PGC-1-/- OIR retinas.
Supp. Fig. 6.-The graph shows the number of tuffs per retinal cross section from OIR wild-type and PGC-1-/- mice. Data are means ±SD (*) p ≤ 0.05 vs. Control.
Supp. Fig. 7.-H&E stain of a retina cross section from an OIR PGC-1-/- mouse with hemorrhages.
Supp. Fig. 8.-Confocal microscope images of isolectin B4 labled retinas from EUK-189 treated or control OIR PGC-1-/- mice (60Xobjective).
Supp. Fig. 9.-qRT-PCR analysis of mRNA of relevant genes in the control of angiogenesisfrom retinas of PGC-1-/- OIR mice. Data are means ±SD (*) p ≤ 0.05 vs. Control.
Supp. Fig. 10.-Left panel, Confocal microscope images of TOMM22 labeled MLEC from wild-type and PGC-1-/-- mice (40X and 60Xobjectives). Right panel, the graph sows the result of image quantitation using Image J software.
Supp. Fig. 11. Representative WB of protein oxidation adducts detected in MLEC from wild-type and PGC-1-/-- as detected with an anti-HNE antibody. the right panel shows the quantification of the 37 kDa band. The bottom panel shows a WB for PGC-1.
References
1. Kuznetsov AV, Troppmair J, Sucher R, Hermann M, Saks V, Margreiter R (2006) Mitochondrial subpopulations and heterogeneity revealed by confocal imaging: possible physiological role? Biochim Biophys Acta 1757 (5-6):686-691. doi:10.1016/j.bbabio.2006.03.014
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