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

Journal: bioRxiv

Article Title: Low-intensity vibration (LIV) cellular mechanotherapy reprograms tumor transcriptomes to suppress cancer-promoting inflammation

doi: 10.64898/2026.04.07.716739

Figure Lengend Snippet: (A) Schematic of the custom-built vibration system, which uses a rigidly mounted gun with adjustable stoppers for tuning vibration amplitude to ensure reproducible mechanical stimulation. (B) Overview of mechanotransduction: extrinsic mechanical forces are converted into biochemical signals that activate downstream pathways, leading to changes in gene expression, protein synthesis, and cell phenotype—thereby, regulating cellular proliferation, differentiation, and apoptosis. (C) Calibration of vibration frequencies at different gear settings: gears 1, 2, and 4 corresponded to 90 Hz, 150 Hz, and 300 Hz, respectively, measured at the plate bottom. Vibration frequency varied inversely with displacement amplitude ( n = 4; p < 0.05). (D) Supernatant analysis of LPS-stimulated PBMCs showed a linear relationship between LPS concentration (0– 10 µg/ml) and IL-6 secretion ( p < 0.05). Maximal PBMC proliferation was observed at 5 μg/ml of LPS ( n = 3). (E) Fold change (FC) of IL-6 cytokine secreted by vibrated PBMCs showed most reduction at the 90 Hz LIV ( n = 3). (F) Time-course of IL-6 secretion from LPS-stimulated PBMCs revealed peak cytokine levels at 48 h ( n = 3). (G) LIV increases the supernatant pH of LPS-stimulated PBMCs in a frequency-dependent manner ( n = 4). (H) LIV did not significantly alter the proliferation of MDA-MB-231 cells compared to static controls ( n = 5; p = 0.374).

Article Snippet: The kits utilized included the human IL-6 ELISA Kit from Elabscience (E-EL-H6156), the IFN-γ ELISA kit from Invitrogen (HHC4021), the CXCL12β ELISA Kit from Invitrogen (EHCXCL12B), the MCP-1 ELISA Kit (BMS281INST), the TNFα ELISA Kit from Invitrogen (KHC3014), and the IL-10 ELISA Kit from Elabscience (EHIL10).

Techniques: Gene Expression, Concentration Assay

Journal: bioRxiv

Article Title: Low-intensity vibration (LIV) cellular mechanotherapy reprograms tumor transcriptomes to suppress cancer-promoting inflammation

doi: 10.64898/2026.04.07.716739

Figure Lengend Snippet:

Article Snippet: The kits utilized included the human IL-6 ELISA Kit from Elabscience (E-EL-H6156), the IFN-γ ELISA kit from Invitrogen (HHC4021), the CXCL12β ELISA Kit from Invitrogen (EHCXCL12B), the MCP-1 ELISA Kit (BMS281INST), the TNFα ELISA Kit from Invitrogen (KHC3014), and the IL-10 ELISA Kit from Elabscience (EHIL10).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: bioRxiv

Article Title: Low-intensity vibration (LIV) cellular mechanotherapy reprograms tumor transcriptomes to suppress cancer-promoting inflammation

doi: 10.64898/2026.04.07.716739

Figure Lengend Snippet: Cytokine concentrations in cell culture supernatants were quantified by ELISA (mean ± SD) and statistical significance was determined by Student’s t-test. (A) IL-6 concentration in vibrated PBMCs was significantly reduced compared to static control (488.26±31.49 pg/ml vs. 573.76± 38.66pg/ml, p = 0.001, n = 7). (B) IFNγ in LIV-treated cells was below the lower limit of detection (LOD) compared to static control concentration of 854.41± 36.01 pg/ml, p < 0.001, n = 5). (C) IL-10 concentration in vibrated PBMCs showed a non-significant increase over control (2.8± 0.83 pg/ml vs. 2.0± 0.70 pg/ml, + 40 %; p = 0.08, n = 5). (D) MCP-1 concentration increased markedly in vibrated PBMCs (4305± 975 pg/ml vs. 2001± 1183 pg/ml, + 115 %; p = 0.02, n = 4). (E) IL-6 in vibrated MDA-MB-231 cells was significantly lower than in controls (109.93± 9.03 pg/ml vs. 171.35± 39.61 pg/ml, − 36 %, p = 0.02, n = 4). (F) TNF-α levels did not differ significantly between vibrated and control MDA-MB-231 cells (24.57± 3.22 pg/ml vs. 28.88± 3.17 pg/ml, − 15 %; p = 0.17, n = 3). (G) CXCL-12β in LIV-treated MDA-MB-231 cells was below the LOD, whereas controls measured at 56.25 ± 39.40 pg/ml ( p = 0.03, n = 4). (H) In co-culture models, IL-6 levels were higher in 2D than in 3D spheroids under LIV (6518± 501 pg/ml vs. 5351± 355 pg/ml, − 22%; p = 0.001, n = 8). (I) IL-6 concentration in vibrated 3D co-cultures was significantly reduced compared with static controls (5351± 355 pg/ml vs. 7104± 392 pg/ml, − 25 %, p < 0.001, n = 8). (J) IL-6 levels in 2D co-cultures did not differ significantly between vibrated and control groups (6518± 501 pg/ml vs. 6887± 483 pg/ml, −5%; p = 0.16, n = 8).

Article Snippet: The kits utilized included the human IL-6 ELISA Kit from Elabscience (E-EL-H6156), the IFN-γ ELISA kit from Invitrogen (HHC4021), the CXCL12β ELISA Kit from Invitrogen (EHCXCL12B), the MCP-1 ELISA Kit (BMS281INST), the TNFα ELISA Kit from Invitrogen (KHC3014), and the IL-10 ELISA Kit from Elabscience (EHIL10).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Concentration Assay, Control, Co-Culture Assay