Fig. 2

Characterisation of SMMHC-HIF-1α^fl/fl mouse line. A Genomic PCR genotyping confirming excision of floxed HIF-1α exon 2 in cortical DNA isolated from tamoxifen treated SMMHC-CreER^T2; HIF-1α^fl/fl mice (LoF) in normoxia or hypoxia. A short 300 bp truncated HIF-1α fragment (HIF-1α^Δ) is produced compared to the 1.1Kbp full length product (HIF-1α^F) obtained in oil-treated controls. B Co-immunostaining with NG-2 or PDGFR-β (green) and Cre recombinase (red) locates Cre recombinase expression specifically to brain pericytes in cortical regions. Scale bar = 50 µm. C Immunoblot of brain cytoplasmic and nuclear fractions confirm tamoxifen-induced nuclear translocation of Cre recombinase in SMMHC-CreER^T2; HIF-1α^fl/fl mice. β-actin was used as loading control. Quantitative RT-PCR expression of HIF-1 targets Glut1 (D) and VEGF (E) in primary pericytes isolated from SMMHC-CreERT2; HIF-1α^fl/fl mice and treated with 2 µM tamoxifen (LoF, loss of function) compared to hypoxic vehicle controls (oil, Ctrl) after 48 h hypoxic exposure (1% O₂). Two-way ANOVA, mean ± SD, n = 4–5. ****p < 0.0001 compared to Hx Ctrl