Endogenous FGF2 and HIF-1α expression in ischemic skin extracts.

<p>A. Representative Western blot analyses of FGF2 and HIF-1α at the indicated post-operative time. The molecular weights of FGF2 mouse isoforms are indicated. B. Levels of endogenous FGF2 mRNA determined by RT-qPCR analysis. Results are expressed relative to the level from control mice and represent mean±SE (n = 5 mice per post-operative time). *P<0.05 vs. control. C. Quantification of total FGF2 detected by Western blot analysis after normalization to GAPDH and to endogenous mRNA level. **P<0.001 vs. control.</p

Crosstalk between FGF2 and HIF-1α.

<p>A. FGF2 and HIF-1α gene silencing using targeted-siRNA. 911 cells were transfected with FGF2 (left) or HIF-1α (right) targeted siRNA (siFGF2 or siHIF-1α, respectively). Protein contents were measured in normoxic (FGF2) and hypoxic (HIF-1α) cells 48 h after transfection. SiC corresponds to a control scrambled siRNA. B. Effect of FGF2 gene silencing on HIF-1α accumulation in 911 cells cultivated in normoxic (N) and hypoxic conditions. C. Effect of HIF-1α gene silencing on FGF2 accumulation in 911 cells cultivated in normoxic (N) and hypoxic conditions.</p

Ischemia induced by a dorsal skin flap model in IRES FGF2-Luc transgenic mice.

<p>A. Representation of FGF2 mRNA and mouse protein isoforms (left panel) and of the bicistronic mRNA expressed by the RFL12 transgenic mice (right panel). This bicistronic cassette expresses, under control of the CMV (Cytomegalovirus) promoter, LucR and LucF reporter genes in a cap- or FGF2 IRES-dependent manner, respectively <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0003078#pone.0003078-Creancier1" target="_blank">[18]</a>. B. Ischemia induction using a skin flap model modified from Ceradini <i>et al </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0003078#pone.0003078-Ceradini1" target="_blank">[26]</a> and representative Laser Doppler analysis performed 18, 48 and 72 hours after surgery. A U-shaped peninsular skin incision was created on the dorsal surface of 8-week old female RFL12 mice. The two vascular pedicles arising from the lateral thoracic arteries were sectioned. To avoid necrotic tissues, the study of gene expression was performed on the proximal part of the skin flap indicated by the white square. The color scale illustrates blood flow variations from maximal (red) to minimal perfusion (dark blue). C. Quantification of laser Doppler analysis. Ctr corresponds to non-operated mice. Results represent mean±SE on at least 3 mice per post-operative time.</p