FIG. 3.

Adipogenic differentiation of mouse adipose-derived stromal cells (mASCs) exposed to various oxygen tensions, with or without HDACi pretreatment. ASCs were expanded under 21%, 10%, and 1% O₂ conditions and were pretreated for 48 h with NaB (1 mM), VPA (1 mM), or vehicle as a control, followed by adipogenic induction. (A) Oil red O staining of intracellular lipid accumulation at 7-day differentiation. Lipid accumulation was observed in 21% and 10% O₂, while little staining was apparent at 1% O₂. HDACi-pretreated mASCs showed substantially reduced lipid at all oxygen tensions. (B) Top row demonstrates quantification of Oil red O staining by leaching and photometric detection. Significantly decreased staining was observed among both NaB- and VPA-pretreated ASCs. Bottom row shows PPAR-γ expression by quantitative real-time polymerase chain reaction. Significantly decreased gene expression in PPAR-γ was again observed among HDACi-pretreated ASCs. Photographs are the scanning view of whole wells and the 10 × microscopical fields that represent one of the triplicate wells; values are normalized to control groups at each oxygen tension; error bars represent one standard deviation; n = 3; *p ≤ 0.05. Color images available online at www.liebertonline.com/ten.