mouse gal3 duoset elisa kit  (R&D Systems)

Journal: Viruses

Article Title: Galectin-3-ITGB1 Signaling Mediates Interleukin 10 Production of Hepatic Conventional Natural Killer Cells in Hepatitis B Virus Transgenic Mice and Correlates with Hepatocellular Carcinoma Progression in Patients

doi: 10.3390/v16050737

Figure Lengend Snippet: High expression levels of Galectin-3 in HBsAg + hepatocytes. Liver and serum samples were collected from 8- to 10-week-old, 4- to 5-month-old and 12- to 13-month-old HBs-Tg mice and control WT B6 mice, respectively. ( A ) mRNA expression levels of Lgals3 in livers were detected by quantitative real-time PCR. ( B ) Serum levels of galectin-3 (Gal-3) protein detected by ELISA. ( C ) Levels of Gal-3 protein in liver tissues detected by ELISA by using liver homogenate. ( D ) Liver samples were collected from 4- to 5-month-old HBs-Tg mice and control WT B6 mice for Gal-3 staining by IHC. Dark-brown dots represent positive staining. Bar = 100 μm. Data are shown as the mean ± SEM. One-way ANOVA test was used. No significant statistical difference is defined as ns. * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: A Galectin-3 mouse ELISA Kit (Invitrogen, Carlsbad, CA, USA) was used to determine the serum and liver homogenate levels of Galectin-3.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Staining

Journal: Viruses

Article Title: Galectin-3-ITGB1 Signaling Mediates Interleukin 10 Production of Hepatic Conventional Natural Killer Cells in Hepatitis B Virus Transgenic Mice and Correlates with Hepatocellular Carcinoma Progression in Patients

doi: 10.3390/v16050737

Figure Lengend Snippet: Galectin-3 induced IL-10 production in hepatic cNK cells and prevented hepatocyte damage in HBs-Tg mice. MNCs were isolated from livers of 4- to 5-month-old HBs-Tg mice and control WT B6 mice. ( A ) IL-10 expression in hepatic cNK cells stimulated by rmGal-3. Liver MNCs (5 × 10 5 ) were stimulated by recombinant mouse Gal-3 (2.5 μg/mL or 5 μg/mL) in 200 μL of 10% FBS 1640 medium for 48 h. Cultured MNCs were harvested and analyzed by flow cytometry. cNK cells (CD3 − NK1.1 + CD49b + CD49a − ) were gated to analyze IL-10 expression. Histograms and MFI are shown. ( B ) Levels of IL-10 protein in culture supernatant. Liver MNCs (5 × 10 5 ) were stimulated by recombinant mouse Gal-3 (50 ng/mL) in 400 μL of 10% FBS 1640 medium for 48 h. Culture supernatant was collected and detected by IL-10 CBA flex set. ( C ) Monensin (2.5 μg/mL) was used to block secretion of IL-10 by cultured NK cells in vitro. Paired Student’s t test was used. * p < 0.05, ** p < 0.01. TD139 (Gal-3-INH) (15 μg/g body weight, once every 24 h for three times) was used. ( D ) Serum levels of IL-10 in HBs-Tg mice after TD139 (Gal-3-INH) treatment detected by ELISA. ( E ) IL-10 expression in liver tissues detected by IHC. Dark-brown dots represent positive staining. Bar = 50 μm. ( F ) Hepatic cNK cells (CD3 − NK1.1 + CD49b + CD49a − ) were gated and analyzed for expression of NKG2D. ( G ) Serum levels of ALT after TD139 (Gal-3-INH) treatment. Data are shown as mean ± SEM. Unpaired Student’s t test was used. * p < 0.05, ** p < 0.01.

Article Snippet: A Galectin-3 mouse ELISA Kit (Invitrogen, Carlsbad, CA, USA) was used to determine the serum and liver homogenate levels of Galectin-3.

Techniques: Isolation, Expressing, Recombinant, Cell Culture, Flow Cytometry, Blocking Assay, In Vitro, Enzyme-linked Immunosorbent Assay, Staining

Journal: Viruses

Article Title: Galectin-3-ITGB1 Signaling Mediates Interleukin 10 Production of Hepatic Conventional Natural Killer Cells in Hepatitis B Virus Transgenic Mice and Correlates with Hepatocellular Carcinoma Progression in Patients

doi: 10.3390/v16050737

Figure Lengend Snippet: Galectin-3 induced IL-10 transcription via ITGB1 signaling in hepatic cNK cells in HBs-Tg mice. MNCs were isolated from livers and spleens of 12- to 13-month-old HBs-Tg mice. LrNK cells (CD3 − NK1.1 + CD49b − CD49a + ) and cNK cells (CD3 − NK1.1 + CD49b + CD49a − ) were gated to analyze expression of ITGB1 (CD29) by flow cytometry analysis. ( A ) Representative histograms of CD29 expression. ( B ) MFI of CD29 expression. ( C ) Binding rate of Gal-3 with hepatic cNK cells compared with splenic cNK cells in HBs-Tg mice. ( D ) Relative mRNA expression levels of IL-10 in hepatic cNK cells. Hepatic cNK cells from the livers of 4- to 5-month-old HBs-Tg mice were purified by MACS. cNK cells (1 × 10 5 ) were plated and then stimulated with recombinant mouse Gal-3 (2.5 μg/mL) for 48 h. Anti-CD29 mAb (30 μg/mL) was used to block interaction between Gal-3 and CD29. Hamster IgG (30 μg/mL) was used as control. Data are shown as mean ± SEM. One-way ANOVA or Student’s t test was used. No significant statistical difference is defined as ns. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: A Galectin-3 mouse ELISA Kit (Invitrogen, Carlsbad, CA, USA) was used to determine the serum and liver homogenate levels of Galectin-3.

Techniques: Isolation, Expressing, Flow Cytometry, Binding Assay, Purification, Recombinant, Blocking Assay

Journal: Viruses

Article Title: Galectin-3-ITGB1 Signaling Mediates Interleukin 10 Production of Hepatic Conventional Natural Killer Cells in Hepatitis B Virus Transgenic Mice and Correlates with Hepatocellular Carcinoma Progression in Patients

doi: 10.3390/v16050737

Figure Lengend Snippet: LGALS3 and ITGB1 expression correlated with poor progression and survival of LIHC patients. Gene Expression Profiling Interactive Analysis (GEPIA2) was used to perform analysis for given sets of TCGA expression data. ( A ) Box plots of LGALS3 gene expression in liver hepatocellular carcinoma (LIHC). TCGA normal and GTEx data were matched. Log2 (TPM + 1) was used for log-scale. * p < 0.05. ( B ) Pathological stage plots of LGALS3 gene expression in LIHC. Use major stages for plotting. Log2 (TPM + 1) was used for log-scale. ( C ) Overall survival analysis of LIHC patients stratified by LGALS3 gene. ( D ) Overall survival analysis of LIHC patients stratified by ITGB1 gene. Median group cutoff was used. Hazards ratio was calculated based on Cox PH Model; 95% confidence interval (CI) was added as a dotted line.

Article Snippet: A Galectin-3 mouse ELISA Kit (Invitrogen, Carlsbad, CA, USA) was used to determine the serum and liver homogenate levels of Galectin-3.

Techniques: Expressing