General characteristics of the older people.

<p>General characteristics of the older people.</p

Flow diagram of this study.

<p>Flow diagram of this study.</p

Adjusted odds ratios for the three exercise types.

<p>Adjusted odds ratios for the three exercise types.</p

Subject characteristics by exercise type.

<p>Subject characteristics by exercise type.</p

General characteristics of the older people.

<p>General characteristics of the older people.</p

Characteristics of subjects by knee osteoarthritis.

<p>Characteristics of subjects by knee osteoarthritis.</p

Hypoxia induces the ERRγ gene expression in hepatoma cell lines.

<p>(<b>A–B</b>), HepG2 cells were seeded in 60-cm² dishes and incubated overnight. Then cells were incubated under hypoxia at indicated time period. The expression of ERRγ was analyzed by Western blot (<b>A</b>) and Q-PCR (<b>B</b>) analysis. (<b>C–D</b>) HepG2 cells were seeded in 60-cm² dishes and incubated overnight and then treated with DFO at indicated concentration and time period. The expression of ERRs was analyzed by Western blot (<b>C</b>) and Q-PCR (<b>D</b>) analysis. ERRγ gene expression was normalized to L32 gene expression, and α or β-tubulin expression. All data are representative of at least three independent experiments. Error bars show ± S.E.M. ^*<i>P</i><0.05, ^**<i>P</i><0.01 by two-tailed Student <i>t</i>-test.</p

Hypoxic activation of HIF-1α directly regulates the transcriptional activity of ERRγ.

<p>(<b>A</b>) HepG2 cells were transfected with hERRγ-Luc. After 24 h of the transfection, HepG2 cells were exposed to hypoxia for indicated time period. Experiments were carried out in triplicate and data are expressed as the fold activation relative to the control. (<b>B–C</b>) HepG2 cells were transfected with hERRγ (−2 kb)-Luc, hERRγ (−1 kb)-Luc, hERRγ (−0.5 kb)-Luc, hERRγ (−0.3 kb)-Luc, hERRγ (HREmt1)-Luc, hERRγ (HREmt2)-Luc, hERRγ (HREmt1+2)-Luc. After 24 h of the transfection, HepG2 cells were exposed to hypoxia for 9 hr and analyzed using luciferase and β-galactosidase assay. Experiments were performed in duplicate and data are expressed as the fold activation relative to the control. (<b>D</b>) ChIP assay: HepG2 cell was exposed to hypoxia for 9 hr. Input represents 10% of purified DNA in each sample. Cell extracts were immunoprecipitated with anti-HIF-1α and purified DNA samples were employed for Q-PCR with primers binding to HRE1 (−1080 to −849) and HRE2 (−508 to −295) and distal site (−1826 to −1586) on the <i>ERRγ</i> gene promoter. All data are representative of at least three independent experiments. Error bars show ± S.E.M. ^***<i>P</i><0.001 by two-tailed Student <i>t</i>-test.</p

ERRγ inverse agonist GSK5182 down-regulates the hypoxia-induced PDK4 expression.

<p>(<b>A–B</b>) HepG2 cells were transfected with hPDK4-Luc. After transfection, the cells were exposed in hypoxia and treated with or without GSK5182. Harvested lysates were utilized for luciferase and β-galactosidase assay (A). Q-PCR was performed using isolated total RNA (B). Experiments were done in triplicate and data are expressed as the fold activation relative to the control. (<b>C</b>) HepG2 cells were seeded in 60-cm² dishes and incubated overnight. The cells were incubated in hypoxia and treated with or without GSK5182. Total protein was harvesed for Western blot analysis using indicated antibodies. (<b>D</b>) HepG2 cells were seeded in 60-cm² dishes and incubated overnight. The cells were treated with or without chemicals (DFO and GSK5182) for 6 hr. Total protein and mRNA were isolated for Western blot assay and RT-PCR and normalized with α or β-tubulin and β-actin. (<b>E</b>) A schematic representation. All data are representative of at least three independent experiments. Error bars show ± S.E.M. ^**<i>P</i><0.01, ^***<i>P</i><0.001 by two-tailed Student <i>t</i>-test.</p

HIF-1α increases the expression of ERR γ.

<p>(<b>A</b>) HepG2 cells were transfected with nonspecific siRNA (NS) or si-HIF-1α, and isolated total protein was analyzed by Western blot. α-tubulin was used as a control. (<b>B</b>) HepG2 cells were transfected with pcDNA3-HIF-1α, pcDNA3-ARNT and hERRγ-Luc, respectively. Experiments were conducted in duplicate and data are expressed as the fold activation relative to the control. (<b>C</b>) HepG2 cells were transfected with pcDNA3-HIF-1α and pcDNA3-ARNT and Q-PCR was performed using isolated total RNA. (<b>D–E</b>) HepG2 cells were transfected with nonspecific siRNA (NS) or si-HIF-1α. After transfection, lysates were utilized for luciferase and β-galactosidase assay (D). Q-PCR was performed using isolated total RNA from HepG2 cells (E). All data are representative of at least three independent experiments. Error bars show ± S.E.M. ^*<i>P</i><0.05, ^***<i>P</i><0.001 by two-tailed Student <i>t</i>-test.</p