Journal: bioRxiv

Article Title: Complex Modulation of IL-6 Signaling by Apelin and Elabela in HTR-8/SVneo Cells Under Cobalt Chloride-Induced Chemical Hypoxia

doi: 10.64898/2026.08.20.746041

Figure Lengend Snippet: The effect of CoCl 2 on the levels of elabela (ELA), apelin, and apelin receptor APJ. The data were obtained using ELA or apelin ELISA assays in HTR-8/SVneo cell homogenates (Panels A and C) or conditioned media (Panels B and D), western blotting (Panel E), or real-time RT-PCR (Panel F) for APJ. Data are mean ± SEM, n=5-6.

Article Snippet: Individual ELISA kits were used for IL-6 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6), sIL-6R (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6sR), and sgp130 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-sgp130) protein quantification in the conditioned media 24 hours after treatment.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Complex Modulation of IL-6 Signaling by Apelin and Elabela in HTR-8/SVneo Cells Under Cobalt Chloride-Induced Chemical Hypoxia

doi: 10.64898/2026.08.20.746041

Figure Lengend Snippet: Elabela (ELA) or apelin treatment decreased IL-6 mRNA in CoCl 2 -treated HTR-8/SVneo cells. Data are mean ± SEM, n=3-6.

Article Snippet: Individual ELISA kits were used for IL-6 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6), sIL-6R (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6sR), and sgp130 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-sgp130) protein quantification in the conditioned media 24 hours after treatment.

Techniques:

Journal: bioRxiv

Article Title: Complex Modulation of IL-6 Signaling by Apelin and Elabela in HTR-8/SVneo Cells Under Cobalt Chloride-Induced Chemical Hypoxia

doi: 10.64898/2026.08.20.746041

Figure Lengend Snippet: Schematic of proposed interactions between IL-6 signaling and apelin and elabela in HTR-8/SVneo trophoblastic cells under CoCl₂-mimicked hypoxia. We hypothesize that apelin/elabela/APJ axis regulates IL-6 signaling via its actions on various components of the IL-6 classical and trans-signaling pathway.

Article Snippet: Individual ELISA kits were used for IL-6 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6), sIL-6R (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6sR), and sgp130 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-sgp130) protein quantification in the conditioned media 24 hours after treatment.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: ASC-Derived Extracellular Vesicles Suppress Macrophage-Driven Inflammatory Amplification and Contractile Activation of Uterine Smooth Muscle Cells

doi: 10.3390/ijms27104273

Figure Lengend Snippet: ASC-derived EVs suppress LPS-induced inflammatory signaling in HUtSMCs. ( A ) HUtSMCs were pretreated with ASC-derived EVs and stimulated with LPS for the indicated time points (5–20 min). Phosphorylation of JNK and ERK1/2 was analyzed by Western blotting. ( B ) mRNA expression levels of TSG6, IL-6, IL-8, MCP-1, and MIP-2 were measured by quantitative RT-PCR at 8–24 h after LPS stimulation and normalized to GAPDH. ( C ) Secretion levels of IL-6, IL-8, and MCP-1 were quantified by ELISA. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: IL-6, IL-8, and MCP-1 levels were measured using ELISA kits, including Human IL-6 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL6-1), Human IL-8 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL8-1), and Human MCP-1 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-MCP1-1), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: ASC-Derived Extracellular Vesicles Suppress Macrophage-Driven Inflammatory Amplification and Contractile Activation of Uterine Smooth Muscle Cells

doi: 10.3390/ijms27104273

Figure Lengend Snippet: ASC-derived EVs modulate TLR4-dependent inflammatory signaling in HUtSMCs. ( A ) HUtSMCs were pretreated with ASC-EVs and/or a TLR4 inhibitor prior to LPS stimulation. Phosphorylation of JNK and ERK1/2 was analyzed by Western blotting. Representative images from three independent experiments are shown. ( B ) mRNA expression levels of IL-6, IL-8, MCP-1, TNF-α, MIP-1α, and MIP-2 were analyzed by quantitative RT-PCR and normalized to GAPDH. Data are presented as mean ± SEM from three independent experiments and are expressed relative to the untreated control. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: IL-6, IL-8, and MCP-1 levels were measured using ELISA kits, including Human IL-6 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL6-1), Human IL-8 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL8-1), and Human MCP-1 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-MCP1-1), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR, Control

Journal: International Journal of Molecular Sciences

Article Title: ASC-Derived Extracellular Vesicles Suppress Macrophage-Driven Inflammatory Amplification and Contractile Activation of Uterine Smooth Muscle Cells

doi: 10.3390/ijms27104273

Figure Lengend Snippet: ASC-derived EVs attenuate macrophage-mediated inflammatory signaling in HUtSMCs. ( A ) HUtSMCs were pretreated with ASC-derived EVs and stimulated with conditioned media (CM) derived from RAW264.7 macrophages treated with LPS (10–100 ng/mL). Cells were harvested after 10 min, and phosphorylation of JNK and ERK1/2 was analyzed by Western blotting. ( B ) mRNA expression levels of IL-6, IL-8, MCP-1, TNF-α, MIP-1α, and MIP-2 were analyzed by quantitative RT-PCR 20 h after CM stimulation. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: IL-6, IL-8, and MCP-1 levels were measured using ELISA kits, including Human IL-6 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL6-1), Human IL-8 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-IL8-1), and Human MCP-1 ELISA Kit (RayBiotech, Peachtree Corners, GA, USA; Cat# ELH-MCP1-1), according to the manufacturer’s instructions.

Techniques: Derivative Assay, Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR

Journal: Cellular and Molecular Immunology

Article Title: Hepatocyte-derived LRG1 primes the liver for metastasis and impairs immunotherapy

doi: 10.1038/s41423-026-01408-9

Figure Lengend Snippet: Tumor-associated inflammation promotes the expression of LRG1 in hepatocytes by IL6/STAT3. A The relative expression of Lrg1 in AML12 cells co-cultured with various mouse tumor cell lines or treated with serum (5%) from CRC model mice. n = 3 independent experiments. Data are means ± SD. B Western blot analysis of the expression LRG1 and β-actin in AML12 cells after the indicated treatments. C Cytokine array analyses of serum from CRC orthotopic mice model (day 7, day 14, day 21 and day 28) and from CRC intrasplenic mice model (day 5, day 10, day 15 and day 21). Quantitative real-time PCR analyses of the expression Lrg1 D and ELISA analyses of media of AML12 E treated with vehicle or recombinant IL6/G-CSF/CXCL13/CCL12/TIMP1. n = 3 independent experiments. Data are means ± SD. F, G qRT-PCR and Western blot analyses of LRG1 in AML12 cells treated with cytokine combinations. n = 3 independent experiments. Data are means ± SD. H, I qRT-PCR and western blot analysis of the expression LRG1 and β-actin in AML12 cells treated with indicated serum and anti-IL6 or tocilizumab. n = 3 independent experiments. Data are means ± SD. J ELISA analyses of serum samples for IL6 from sham-group, CRC orthotopic model at day 21(PMN) and CRC intrasplenic model at day 21(Met) in BALB/c. n = 4. Data are means ± SD. K Correlation between serological IL6 and LRG1 levels of CRC patients is shown using Pearson’s correlation analysis. Dots represent individual samples. n = 161. L, M Schematic of the experimental design L . Representative images of liver metastases M in each group ( n = 6 in Lrg1 (+/+)Hep-HTVi-Ctrl group and Lrg1 (+/+)Hep-HTVi-IL6 group, n = 5 in Lrg1 (Δ/Δ)Hep-HTVi-Ctrl group and Lrg1 (Δ/Δ)Hep-HTVi-IL6 group). Scale bars, 1 cm. N Schematic of the experimental design. Representative bioluminescence images and analyses of liver metastases in CRC mouse model treated with anti-IL6 or IgG. O Schematic of the experimental design. P ELISA analyses of samples for IL6 levels from liver interstitial fluid, peripheral blood and tumor interstitial fluid of mice from different groups as indicated. n = 4. Data are means ± SD. Q ELISA analyses of samples for LRG1 levels from liver interstitial fluid and peripheral blood of mice from different groups as indicated. n = 4. Data are means ± SD. R Western blot analyses of LRG1 and β-actin in hepatocyte of mice from different groups as indicated. Statistical significance was determined using two-tailed unpaired Student’s t test A, D–F, H, and N–P

Article Snippet: ELISA kits were used to measure the levels of human LRG1 (Ray Bio), human IL6 (Boster, EK0410), mouse LRG1 (ELK Biotechnology), and mouse IL6 (Boster, EK0411) in cell culture supernatants or serum samples according to the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Recombinant, Quantitative RT-PCR, Two Tailed Test