HIF-1α is required for disturbed flow-induced metabolic reprogramming in human and porcine vascular endothelium - PubMed (original) (raw)
(A) HAECs were treated with either UF or DF for 24 hr prior to lysis, RNA purification, and qRT-PCR for IL8, VCAM1, and CCL2. IL8, VCAM1, and CCL2 are all increased under DF (n = 4). (B) HAECs were treated with either non-targeting siRNA (SC) or siRNA targeting HIF-1α (si_HIF-1α_) for 24 hr before subjected to DF for 48 hr prior to lysis, RNA purification, and qRT-PCR for IL8, VCAM1, and CCL2. IL8, VCAM1, and CCL2 are all reduced under HIF-1α knockdown (n = 4). (C) HAECs were transfected with either a blank or with stabilized HIF-1α mRNA transcript for 6 hr prior to lysis, RNA purification, and qRT-PCR for IL8, VCAM1, and CCL2. IL8, VCAM1, and CCL2 are all increased under HIF-1α transcription delivery (n = 4). (D) HAECs were treated with either non-targeting siRNA (SC) or siRNA targeting NOX4 (si_NOX4_) for 24 hr before subjected to DF for 48 hr prior to lysis, RNA purification, and qRT-PCR for IL8, VCAM1, and CCL2. IL8, VCAM1, and CCL2 are all reduced under NOX4 knockdown (n = 4). (E) HAECs were treated with either non-targeting siRNA (SC) or siRNA targeting SLC2A1 (si_SLC2A1_) for 24 hr before subjected to DF for 48 hr prior to lysis, RNA purification, and qRT-PCR for IL8, VCAM1, and CCL2. IL8, VCAM1, and CCL2 are all reduced under SLC2A1 knockdown (n = 4). (F) HAECs were treated with either non-targeting siRNA (SC) or siRNA targeting PDK1 (si_PDK1_) for 24 hr before subjected to DF for 48 hr prior to lysis, RNA purification, and qRT-PCR for IL8, VCAM1, and CCL2. IL8, VCAM1, and CCL2 are all reduced under PDK1 knockdown. (G) HAECs were treated with either UF or DF for 24 hr prior to lysis and Western Blotting (n = 4). (H) si_HIF-1α_ treated HAECs were subjected to DF for 48 hr prior to lysis. si_HIF-1α_ significantly reduces phospho-p65 (n = 4). (I) HAECs were treated with 10 μM of NBD for 1 hr prior to HIF-1α overexpression for 6 hr prior to RNA purification and qRT-PCR (n = 4). *p<0.05; **p<0.005; ***p<0.0005 as determined by Student’s t-test. Data represent mean ± SEM. DOI: