Journal: Journal of Lipid Research

Article Title: KLF7 induced ADRB3-dependent IL-6 production in brown adipocytes during stress

doi: 10.1016/j.jlr.2026.100981

Figure Lengend Snippet: Stress induces KLF7 and IL-6 in BAT by activating ADRB3. A: Immunoblotting for p-CREB, CREB, KLF7, and IL-6 protein levels in the BAT of mice 4 h after retro-orbital bleeding. Quantification of band intensity was performed by densitometric analysis, normalized to β-Tubulin. Band intensities for p-CREB were normalized to total CREB. B: mRNA levels of the Il-6 gene in the BAT of mice. C: Plasma IL-6 levels in mice after bleeding and injection of the ADRB3 antagonist SR59203A. D: Immunoblotting for p-PKA substrates in the BAT of mice. E: Klf7 mRNA expression in the BAT of mice after bleeding. F: Blood glucose levels of mice after bleeding. G: PTT was performed in mice 4 h after bleeding. AUC, area under the curve. H: Gluconeogenesis-associated gene expression in the liver of mice after bleeding. I: Protein levels of the p-PKA substrate in mice 2 h post injection of the ADRB3 agonist CL316,243 and the ADRB3 antagonist SR59203A. J: Immunoblotting for p-CREB, CREB, KLF7, and IL-6. K: Quantification of KLF7 and IL-6 protein expression levels by grayscale value analysis. L and M: mRNA levels of Klf7 and Il-6 in mice 2 h after administration of CL316,243 and SR59203A. N: Circulating IL-6 levels in mice postinjection. O: mRNA levels of gluconeogenesis-associated genes in the liver. n = 5 per group. Western blot quantification represents densitometric analysis from three independent biological samples. The data are presented as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. BAT, brown adipose tissue; IL-6, interleukin-6; KLF7, Krüppel-like factor 7; PKA, protein kinase A; CREB, cAMP response element-binding protein; PTT, pyruvate tolerance test.

Article Snippet: IL-6 levels in serum and cell culture medium were measured using a mouse IL-6 ELISA kit (Proteintech, KE10007, China) following the manufacturer's instructions.

Techniques: Western Blot, Clinical Proteomics, Injection, Expressing, Gene Expression, Binding Assay

Journal: Journal of Lipid Research

Article Title: KLF7 induced ADRB3-dependent IL-6 production in brown adipocytes during stress

doi: 10.1016/j.jlr.2026.100981

Figure Lengend Snippet: ADRB3 activation increases KLF7 and IL-6 expression in brown adipocytes. A: Phosphorylation of the PKA substrate in primary brown adipocytes treated with 5 μM CL316,243, normalized to β-Tubulin. B: CREB phosphorylation in brown adipocytes, normalized to total CREB. C: Immunoblotting and densitometric quantification of KLF7 and IL-6 protein levels in CL316,243-treated primary brown adipocytes. D: mRNA expression of Klf7 and Il-6 . E: IL-6 levels in the medium. F: Gluconeogenesis-associated gene expression in Hepa 1–6 cells cultured with conditioned medium from primary brown adipocytes treated with CL316243 . Data are shown as mean ± SD from three independent biological experiments, each performed using independently isolated and differentiated primary brown adipocytes. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. IL-6, interleukin-6; KLF7, Krüppel-like factor 7; PKA, protein kinase A; CREB, cAMP response element-binding protein.

Article Snippet: IL-6 levels in serum and cell culture medium were measured using a mouse IL-6 ELISA kit (Proteintech, KE10007, China) following the manufacturer's instructions.

Techniques: Activation Assay, Expressing, Phospho-proteomics, Western Blot, Gene Expression, Cell Culture, Isolation, Binding Assay

Journal: Journal of Lipid Research

Article Title: KLF7 induced ADRB3-dependent IL-6 production in brown adipocytes during stress

doi: 10.1016/j.jlr.2026.100981

Figure Lengend Snippet: ADRB3-induced IL-6 expression requires Klf7 in adipocytes during stress. A: Representative genotyping results for WT and Klf7 AKO mice. B: Representative western blots and densitometric quantification showing KLF7 levels in the BAT of WT and Klf7 AKO mice. C: Klf7 mRNA levels in the BAT of WT and Klf7 AKO mice. D: The levels of IL-6 in Klf7 AKO or WT mice after bleeding. E: mRNA expression of IL-6 in BAT. F: Circulating IL-6 levels. G: Gluconeogenic gene expression in the livers of WT and Klf7 AKO mice. H: PTT was performed in mice after bleeding. I: Blood glucose levels in mice after bleeding (n = 5 per group). J: Protein levels of IL-6 in mice 2 h after administration of CL316,243. K: mRNA expression in the BAT of mice after treated with 5 mg/kg CL316,243. L: Circulating IL-6 levels. M: Gluconeogenesis-associated gene expression in the livers of WT and Klf7 AKO mice. NT, no treated. n = 5 per group. Western blot quantification represents densitometric analysis from three independent biological samples. The data are presented as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. BAT, brown adipose tissue; IL-6, interleukin-6; KLF7, Krüppel-like factor 7; Klf7 AKO, Klf7 -adipocyte knockout; PTT, pyruvate tolerance test.

Article Snippet: IL-6 levels in serum and cell culture medium were measured using a mouse IL-6 ELISA kit (Proteintech, KE10007, China) following the manufacturer's instructions.

Techniques: Expressing, Western Blot, Gene Expression, Knock-Out

Journal: Journal of Lipid Research

Article Title: KLF7 induced ADRB3-dependent IL-6 production in brown adipocytes during stress

doi: 10.1016/j.jlr.2026.100981

Figure Lengend Snippet: Stress-induced upregulation of KLF7 and IL-6 in BAT is mediated by PKA activation. A: Representative western blots and densitometric quantification of PKA substrate phosphorylation after bleeding and administration of the PKA inhibitor H89. B: p-CREB, CREB, KLF7, and IL-6 protein levels in BAT. C: Klf7 and Il-6 mRNA levels in the BAT of mice 4 h after retro-orbital bleeding and the injection of the PKA inhibitor H89. D: Plasma IL-6 levels. E: Gluconeogenesis-associated gene expression in the liver of mice after bleeding. F: PTT was performed in mice 4 h after bleeding. G: Blood glucose levels in mice after bleeding. n = 5 per group. H and I: PKA substrate and CREB phosphorylation in the BAT of mice after the injection of CL316,243 and H89. J and K: mRNA and protein levels of Klf7 and Il-6 . L: Plasma levels of IL-6. M: mRNA levels of gluconeogenic genes in the liver. n = 5 per group. Western blot quantification represents densitometric analysis from three independent biological samples. The data are presented as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. BAT, brown adipose tissue; IL-6, interleukin-6; KLF7, Krüppel-like factor 7; PKA, protein kinase A; CREB, cAMP response element-binding protein; PTT, pyruvate tolerance test.

Article Snippet: IL-6 levels in serum and cell culture medium were measured using a mouse IL-6 ELISA kit (Proteintech, KE10007, China) following the manufacturer's instructions.

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Injection, Clinical Proteomics, Gene Expression, Binding Assay

Journal: Journal of Lipid Research

Article Title: KLF7 induced ADRB3-dependent IL-6 production in brown adipocytes during stress

doi: 10.1016/j.jlr.2026.100981

Figure Lengend Snippet: PKA mediates the upregulation of KLF7 and IL-6 expression induced by ADRB3 activation and cAMP elevation in brown adipocytes. A: p-PKA substrate levels in primary brown adipocytes treated with CL316,243 and H89, normalized to β-tubulin. B: p-CREB, CREB, KLF7, and IL-6 protein levels in H89-pretreated primary brown adipocytes, normalized to β-tubulin. Band intensities for p-CREB were normalized to total CREB. C and D: K lf 7 and I l -6 mRNA expression. E: IL-6 levels in the medium of primary brown adipocytes treated with the indicated compounds (5 μM CL316243 or 10 μM H89). F: mRNA levels of gluconeogenesis-associated genes in Hepa 1–6 cells cultured with conditioned medium from primary brown adipocytes treated with the indicated compounds (5 μM CL316243 , 10 μM H89, or 2 μg/ml anti-IL-6 mAb). G: p-PKA substrate levels in primary brown adipocytes treated with 10 μM forskolin and 10 μM H89. H: p-CREB, CREB, KLF7, and IL-6 protein expression in primary brown adipocytes. I: mRNA expression of Klf7 in forskolin- and H89-treated primary brown adipocytes. J: Il-6 mRNA levels in primary brown adipocytes. K: IL-6 levels in the medium of brown adipocytes. L: Gluconeogenesis-associated gene expression levels in Hepa 1–6 cells cultured with conditioned medium from primary brown adipocytes treated with the indicated compounds (10 μM forskolin, 10 μM H89, or 2 μg/ml anti-IL-6 mAb). Data are shown as mean ± SD from three independent biological experiments, each performed using independently isolated and differentiated primary brown adipocytes. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. IL-6, interleukin-6; KLF7, Krüppel-like factor 7; PKA, protein kinase A; CREB, cAMP response element-binding protein.

Article Snippet: IL-6 levels in serum and cell culture medium were measured using a mouse IL-6 ELISA kit (Proteintech, KE10007, China) following the manufacturer's instructions.

Techniques: Expressing, Activation Assay, Cell Culture, Gene Expression, Isolation, Binding Assay

Journal: Journal of Lipid Research

Article Title: KLF7 induced ADRB3-dependent IL-6 production in brown adipocytes during stress

doi: 10.1016/j.jlr.2026.100981

Figure Lengend Snippet: CREB mediates ADRB3-induced KLF7 and IL-6 expression in brown adipocytes and transcriptionally activates KLF7. A: Immunoblotting for p-CREB, CREB, KLF7, and IL-6 in 666-15-pretreated primary brown adipocytes, normalized to β-tubulin. Band intensities for p-CREB were normalized to total CREB. B: Klf7 mRNA levels. C: Il-6 mRNA levels. D: IL-6 levels in the medium of primary brown adipocytes treated with the indicated compounds (5 μM CL316,243 and 10 μM 666-15). E: Binding site of CREB to KLF7 as predicted by the JASPAR database. F: Schematic of KLF7 fluorescent plasmids. G: KLF7 fluorescence activity values in CREB-overexpressing HEK-293T cells. H: ChIP assay performed in CREB-overexpressing HEK-293T cells. Data are shown as mean ± SD from three independent biological experiments performed using independently prepared primary brown adipocytes or HEK-293T cells. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. IL-6, interleukin-6; KLF7, Krüppel-like factor 7; CREB, cAMP response element-binding protein; ChIP, chromatin immunoprecipitation.

Article Snippet: IL-6 levels in serum and cell culture medium were measured using a mouse IL-6 ELISA kit (Proteintech, KE10007, China) following the manufacturer's instructions.

Techniques: Expressing, Western Blot, Binding Assay, Fluorescence, Activity Assay, Chromatin Immunoprecipitation

Journal: Cancer Research

Article Title: Slit2 Inhibits Breast Cancer Metastasis by Activating M1-Like Phagocytic and Antifibrotic Macrophages

doi: 10.1158/0008-5472.can-20-3909

Figure Lengend Snippet: Figure 5. Slit2 reduces the expression of IL6, thereby inhibiting TAMs activity. A, Total RNA of CD11bþ cells sorted from rSlit2-N PBS-treated MMTV-PyMT tumorswas subjected to gene expression analysis using NanoString technology. The heatmap shows differentially expressed top genes. B, Total RNA was isolated from MDA-MB-231 cells overexpressing Slit2 (231-Slit2) or vector control (231-vec) and analyzed for gene expression using microarray technology. The heatmap shows differentially expressed top genes. C, 231-Slit2 or 231-Vec cells CM was subjected to cytokine array analysis. The representative image shows differentially expressed molecules. D, The graph shows levels of IL6 in CM derived from 231-Sli2 or Vec CM detected by ELISA. E, Expression of Robo1 in MDA-MB-231 cells transduced with lentivirus expressing siRNA specific to Robo1 (si-Robo1) or control (si-ctrl) by Western blot. F, b-actin was used as loading control. MDA-MB-231 parental control, si-Robo1, or si-ctrl from E was treated with rSlit2-N or PBS and phosphorylation at p65 of NFkB (p-NFkB) or total NFkB (t-NFkB) was analyzed. G, Graph showing levels of IL6 in si-Robo1 or si-ctrl MDA-MB-231 cells treated with rSlit2-N or PBS detected by ELISA (n ¼ 3 each group). H, BMDMs were pretreated with serum-free CM (CTRL), or 231-Slit2 CM or 231-Vec CM or 231-Vec CM preincubated with IL6 nAb (231-Vec þ IL6nAb) or IL6 nAb. After 2 hours, pH-Rhodo–labeled S. Aureus particles were added to the cells and recombinant IL6 was added to the 231-Slit2 CM cells (231-Slit2-rIL6). The kinetics of S. Aureus phagocytosis was analyzed using a fluorescence plate reader at every 15 minutes up to 4 hours. , P < 0.05; , P < 0.01; , P < 0.001 using Student t test.

Article Snippet: The cells were pretreated with mouse rSlit2-N (100 ng/mL) or IL6 neutralization antibody (BioXCell# BE0046) for 2 hours.

Techniques: Expressing, Activity Assay, Gene Expression, Isolation, Plasmid Preparation, Control, Microarray, Derivative Assay, Enzyme-linked Immunosorbent Assay, Transduction, Western Blot, Phospho-proteomics, Labeling, Recombinant