Effects of PI3K inhibitors on the baicalein–activated HIF1α and Epo/VEGF gene transcription in neurons.

<p>Neurons were treated with 3.5 μM BA with or without 1 h pretreatment with the pan PI3K inhibitor LY294002 (LY, 10 μM), PI3Kα/β inhibitor (PI3K α inhibitor-2, 50 nM), or PI3Kγ inhibitor (CAY10505, 200 nM). (A) Cells were harvested at 30 min after the BA treatment for Western blotting of pAkt and Akt. (B) Dual-luciferase activity assay of pHRE_<i>EPO</i>-Luc expression in cells 24 h after the treatments. (C) qRT-PCR of Epo and VEGF mRNA expression in cells 24 h after the treatments. (D) Anti-HIF1α-based ChIP assay was performed at 0, 1, 3 h after the BA treatment to analyze the HIF1α-associated rat <i>Epo</i> promoter fragment (<i>Epo-p</i>), HRE-containing <i>Epo</i> enhancer fragment (<i>Epo-e</i>) and HRE-containing <i>Vegf</i> promoter fragment (<i>Vegf-p</i>) by PCR (left panel) and qPCR (right panel). *p<0.05, **p<0.01, ***p<0.001 versus control; ^# p<0.05, ^{# #} p<0.01, ^{# # #} p<0.001 versus the BA-treated group (n=3).