Correlation with Log Fstl1 levels.

<p>Correlation with Log Fstl1 levels.</p

Correlation between plasma Fstl1 levels and clinical parameters.

<p>Correlation of plasma Log Fstl1 levels with Log fasting immune-reactive insulin (FIRI), Log high sensitive CRP (hsCRP) and derivatives of reactive oxidative metabolites (dROMs) was analyzed.</p

Correlation with dROMs levels.

<p>Correlation with dROMs levels.</p

Correlation with Log hsCRP levels.

<p>Correlation with Log hsCRP levels.</p

Clinical characteristics of all subjects.

<p>Clinical characteristics of all subjects.</p

Ablation of KLF15 by siRNA reduces adipolin expression in adipocytes.

<p>KLF15, adipolin (APL) and adiponectin (APN) mRNA levels were determined by quantitative RT-PCR method. <b>A</b>, KLF15 mRNA levels in 3T3-L1 adipocytes at 48 h after transfection with siRNA targeting KLF15 (si-KLF15) (20 nM) or non-targeting control siRNA (si-Control) (20 nM). N = 3 in each group. <b>B</b>, mRNA levels of APL and APN in 3T3-L1 adipocytes transfected with si-KLF15 (20 nM) or si-Control (20 nM). N = 3 in each group.</p

Overexpression of KLF15 rescues the reduction of adipolin expression caused by TNFα.

<p>Quantitative RT-PCR method was used for measurement of mRNA levels. <b>A</b>, Adipolin mRNA levels treated with adenovirus expressing KLF9 (Ad-KLF9), KLF15 (Ad-KLF15) or β-galactosidase (Ad-β-gal) at 150 moi for 24 h in 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated with TNFα (10 ng/ml) or vehicle for 24 h. N = 3 in each group. <b>B</b>, Adiponectin mRNA levels treated with Ad-KLF15 or Ad-β-gal at 150 moi for 24 h in 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated with TNFα (10 ng/ml) or vehicle for 24 h. N = 3 in each group.</p

Expression of KLF15 augments the promoter activity of adipolin.

<p><b>A and B</b>, Effect of KLF15 and KLF9 on the promoter activity of adipolin. Protein levels of KLF15 (A) and KLF9 (B) in HEK293 cells transfected with pShuttle vector expressing KLF15 tagged with FLAG, KLF9 tagged with FLAG or empty vector (MOCK). Expression of KLF15 and KLF9 was evaluated by Western blot analyses using anti-FLAG antibody. HEK293 cells were transfected with pShuttle vector expressing KLF15, KLF9 or MOCK, along with pGL3-basic vectors containing adipolin promoter region (−66/−1 or −111/−1) or empty pGL3 vector in the presence of pRL-SV40. Promoter activity was assessed by luciferase reporter assay. Results are normalized relative to the values of empty pShuttle vectors (MOCK). N = 6 in each group. <b>C</b>, Luciferase assay for determination of adipolin promoter activity in 3T3-L1 adipocytes. 3T3-L1 adipocytes were transfected with pGL3-basic vectors containing adipolin promoter (−66/−1 or −111/−1) or empty pGL3 vector in the presence of pRL-SV40. N = 6 in each group.</p

Inhibition of JNK restores TNFα-mediated reduction of adipolin expression in 3T3-L1 adipocytes.

<p>The mRNA levels of adipolin (APL) and KLF15 were measured by quantitative RT-PCR method. <b>A and B</b>, Effect of JNK inhibitor on mRNA levels of APL (A) and KLF15 (B) in adipocytes. 3T3-L1 adipocytes were cultured in the presence or absence of JNK inhibitor, SP600125 (10 µM) for 1 h followed by treatment with TNFα (10 ng/ml) or vehicle for 24 h. N = 3 in each group. <b>C</b>, Effect of p38 MAPK inhibitor on APL mRNA levels in adipocytes. 3T3-L1 adipocytes were treated with or without p38 MAPK inhibitor, SB203580 (10 µM) for 1 h followed by stimulation with TNFα (10 ng/ml) or vehicle for 24 h. N = 3 in each group.</p

KLF9 and KLF15 are down-regulated in adipose tissue of obese mice and TNFα-treated cultured adipocytes.

<p>Quantitative RT-PCR method was used for measurement of mRNA levels. <b>A</b>, mRNA levels of KLF3, KLF9, KLF10, KLF13, KLF15 and adipolin (APL) in adipose tissues of control lean and high fat diet-induced obese (DIO) mice. N = 3–4 in each group. <b>B</b>, mRNA levels of KLF3, KLF9, KLF10, KLF13, KLF15 and adipolin (APL) in 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated with TNF-α (10 ng/ml) or vehicle for 24 h. N = 3 in each group.</p