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

Journal: Journal of Clinical Immunology

Article Title: Mice with Asthma Are More Resistant to Influenza Virus Infection and NK Cells Activated by the Induction of Asthma Have Potentially Protective Effects

doi: 10.1007/s10875-011-9619-2

Figure Lengend Snippet: Asthmatic model mice exhibit increased OVA-specific IgE in their serum and histopathological changes characterized by allergic asthma. C57BL/6 mice were intraperitoneally sensitized with OVA or PBS as a control every other day for 2 weeks, then allowed to rest for 3 weeks. The OVA-sensitized mice or control mice were then intranasally challenged with OVA or PBS on days −3, −2 and −1, respectively. Bloods, trachea tissues, and lungs were collected from the mice on day 0. a The OVA-specific IgE in the serum was measured by an ELISA assay. b The trachea tissues ( upper panels ) and lungs ( bottom panels ) were stained with HE. Six mice were used in each group and similar observations were obtained from each mouse

Article Snippet: The cytokine concentrations in the BALF supernatants were determined using a mouse Interferon Alpha ELISA Kit (PBL Biomedical Laboratories, NJ, USA) and a mouse IL-12p40 ELISA development kit, mouse IFN-γ ELISA development kit, mouse IL-6 ELISA development kit, mouse TNF-α ELISA development kit, mouse IL-4 ELISA development kit, mouse IL-13 ELISA development kit, or a mouse IL-22 ELISA development kit (R&D Systems, MN, USA) according to the manufacturer’s instructions, as described previously [ ].

Techniques: Control, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Clinical Immunology

Article Title: Mice with Asthma Are More Resistant to Influenza Virus Infection and NK Cells Activated by the Induction of Asthma Have Potentially Protective Effects

doi: 10.1007/s10875-011-9619-2

Figure Lengend Snippet: Differences were observed in the cytokine production between asthmatic mice and control mice during influenza virus infection. C57BL/6 mice were sensitized and challenged with OVA or PBS. Subsequently, the mice were infected with 100 pfu of the influenza virus. BALF samples from the mice were collected on day 0 before viral infection and on days 2, 4, and 6 after influenza virus infection, and the cytokine concentrations in the BALF supernatants were measured by ELISA assays. Six mice were used in each group. These results are representative of two independent experiments. * p < 0.05 compared to the cytokine production of control mice. These results are representative of two independent experiments

Article Snippet: The cytokine concentrations in the BALF supernatants were determined using a mouse Interferon Alpha ELISA Kit (PBL Biomedical Laboratories, NJ, USA) and a mouse IL-12p40 ELISA development kit, mouse IFN-γ ELISA development kit, mouse IL-6 ELISA development kit, mouse TNF-α ELISA development kit, mouse IL-4 ELISA development kit, mouse IL-13 ELISA development kit, or a mouse IL-22 ELISA development kit (R&D Systems, MN, USA) according to the manufacturer’s instructions, as described previously [ ].

Techniques: Control, Virus, Infection, Enzyme-linked Immunosorbent Assay

Journal: BMB Reports

Article Title: Effects of α-lipoic acid on LPS-induced neuroinflammation and NLRP3 inflammasome activation through the regulation of BV-2 microglial cells activation

doi: 10.5483/BMBRep.2019.52.10.026

Figure Lengend Snippet: α-LA inhibits the expression of pro-inflammatory cytokines in LPS-treated BV-2 microglial cells. (A) Effects of α-LA on cell viability. BV-2 microglial cells were incubated with LPS (1 μg/ml) for 30 min followed by treatment with the indicated concentrations of α-LA for 24 h. Thereafter, cell viability was assessed through the MTT assay. (B and C) BV-2 microglial cells were treated with LPS (1 μg/ml) for 30 min followed by treatment with the indicated concentrations of α-LA at the suggested times. The cell-free conditioned culture medium was collected and analyzed with ELISA for TNF-α, IL-6. Data from three independent experiments are presented as means ± S.D. *≤ 0.05, **< 0.01, ***< 0.001 and are related to both LPS-induced cells and α-LA treated cells.

Article Snippet: Both TNF-α and IL-6 were quantitatively measured through an enzyme-linked immunosorbent assay (ELISA) using the mouse TNF-α and IL-6 DuoSet ELISA kit (R&D systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.

Techniques: Expressing, Incubation, MTT Assay, Enzyme-linked Immunosorbent Assay