Journal:

Article Title: Vascular MAD s: Two novel MAD -related genes selectively inducible by flow in human vascular endothelium

doi:

Figure Lengend Snippet: Smad6 and Smad7 are regulated genes in vascular endothelium. (A) Northern analysis of RNA from cultured confluent HUVEC monolayers exposed to static (no-flow) conditions, 24 hr of steady LSS (10 dyn/cm2), TSS (10 dyn/cm2), or rhIL-1β (10 units/ml; Biogen) and probed with cDNA for Smad6 or Smad7, or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to control for RNA loading. (B) Northern analysis of RNA from cultured HUVEC treated for 24 hr with tumor necrosis factor α (TNF-α, 200 units/ml; Biogen), interferon-γ (IFN-γ, 500 units/ml; Genzyme), or TGF-β1 (10 ng/ml; Genzyme). All cytokines were of human origin. Bumetanide-sensitive cotransporter 2 (BSC2) is an ion cotransporter that is transcriptionally up-regulated by TNF-α in HUVEC (23). (C) Northern analysis of Smad2 and Smad4 RNA in HUVEC demonstrating that their levels are not significantly altered by 24 hr of steady LSS. (D) Nuclear runoff analysis of HUVEC exposed to static conditions (control), LSS, TSS, or rhIL-1β for 3 hr. The genes encoding Smad6 and Smad7 demon strate a significant increase in their rates of transcription only in response to the LSS stimulus. The cyclooxygenase 1 (Cox-1) and tubulin genes do not demonstrate significant change, and transcription of the intercellular adhesion molecule 1 (ICAM-1) gene is up-regulated by both the LSS and rhIL-1β stimuli as previously reported (2). Rsv-CAT is a negative control plasmid containing no endothelial genes.

Article Snippet: Neither Smad2 nor Smad4 , two other human MAD genes, is up-regulated by steady LSS in cultured HUVEC (Fig. C ). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 2 caption a7 Smad6 and Smad7 are regulated genes in vascular endothelium. ( A ) Northern analysis of RNA from cultured confluent HUVEC monolayers exposed to static (no-flow) conditions, 24 hr of steady LSS (10 dyn/cm 2 ), TSS (10 dyn/cm 2 ), or rhIL-1β (10 units/ml; Biogen) and probed with cDNA for Smad6 or Smad7, or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to control for RNA loading. ( B ) Northern analysis of RNA from cultured HUVEC treated for 24 hr with tumor necrosis factor α (TNF-α, 200 units/ml; Biogen), interferon-γ (IFN-γ, 500 units/ml; Genzyme), or TGF-β1 (10 ng/ml; Genzyme).

Techniques: Northern Blot, Cell Culture, Negative Control, Plasmid Preparation