Journal: International journal of cancer

Article Title: Serum haptoglobin as a novel molecular biomarker predicting colorectal cancer hepatic metastasis.

doi: 10.1002/ijc.29993

Figure Lengend Snippet: Figure 1. Elevated HP levels in colorectal cancer patients. (a) Serum levels of HP in healthy controls and colorectal cancer patients. (b) The serum levels of HP in healthy controls (n 5 152) and colorectal cancer patients [M0, M1(S), and M1(H), n 5 250] were measured by ELISA. (c) Kaplan–Meier survival curve showed correlation between HP serum level and overall survival. (*, p<0.05) [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Article Snippet: Measurement of HP level in human blood samples A commercially available ELISA kit Human Haptoglobin ELISA Kit(Assaypro EH1003-1)was used to measure HP level.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Science Advances

Article Title: Mitochondrial respiratory complex III sustains IL-10 production in activated macrophages and promotes tumor-mediated immune evasion

doi: 10.1126/sciadv.adq7307

Figure Lengend Snippet: ( A to D ) BMDMs were pretreated with DMSO or S3QEL 1.2 (0.1 to 10 μM) for 3 hours before LPS (100 ng/ml) stimulation for 4 hours, and cell lysates and supernatants were harvested. (A) and (B) Quantification of IL-10 mRNA (A) and released protein (B) by RT-qPCR, relative to the Rps18 housekeeping gene, and ELISA. (C) and (D) Quantification of TNF- α mRNA (C) and released protein (D) by RT-qPCR, relative to the Rps18 housekeeping gene, and ELISA. FC, fold change. ( E to H ) BMDMs were pretreated with DMSO and MYX (500 nM) for 3 hours before LPS (100 ng/ml) stimulation for 4 hours, and cell lysates and supernatants were harvested. (E) and (F) Quantification of IL-10 mRNA (E) and released protein (F) by RT-qPCR, relative to the Rps18 housekeeping gene, and ELISA. (G) and (H) Quantification of TNF- α mRNA (G) and released protein (H) by RT-qPCR, relative to the Rps18 housekeeping gene, and ELISA. ( I ) Schematic diagram of in vivo procedure and sample collection. Mice were intraperitoneally (ip) injected with PBS or S3QEL 1.2 (1 mg/kg) for 2 hours, followed by LPS (2.5 mg/kg) for 2 hours. Blood (then serum) was collected. hrs, hours. ( J ) Quantification of IL-10 by ELISA in the serum. Data from (A) to (H) are expressed as means ± SEM for n = 5 to 9 from three independent experiments. Data from (J) are expressed as means ± SEM ( n = 10 per group within two independent in vivo experiments). P values were calculated using one-way ANOVA for multiple comparisons. Differences were considered statistically significant at * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: For in vivo samples, a Quantikine ELISA kit for mouse IL-10 (R&D Systems) was used according to the manufacturer’s instructions with appropriately diluted serum added to each plate.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, In Vivo, Injection

Journal: Science Advances

Article Title: Mitochondrial respiratory complex III sustains IL-10 production in activated macrophages and promotes tumor-mediated immune evasion

doi: 10.1126/sciadv.adq7307

Figure Lengend Snippet: ( A ) BMDMs were pretreated with DMSO, MYX (500 nM), or S3QEL 1.2 (10 μM) for 3 hours, stimulated with CpG (1 μg/ml) for 4 hours, and cell lysates and supernatants were harvested. Quantification of IL-10 mRNA by RT-qPCR, relative to the Rps18 housekeeping gene, and of IL-10 levels by ELISA. ( B ) Schematic diagram of the melanoma in vivo model. Mice were challenged with 5 × 10 6 B16F10 cells subcutaneously and administered with PBS, S3QEL 1.2 (1 mg/kg), and/or CpG (2.5 mg/kg). Tumors were measured daily. D, day. ( C ) Mean initial tumor growth curve relative to tumor volume from day −1 to day 8. ( D ) Kaplan-Meier survival graph up to day 65 (end of experiment). ( E ) Tumor growth rate curve relative to tumor area from day 0 to day 17. ( F to H ) Immune cells infiltrating the tumor were analyzed by flow cytometry. Cells expressing the surface markers Ly6C, F4/80, and CD11b showing the percentage of MHCII + macrophages and mean fluorescence intensity (MFI) of MHCII (F), the percentage of CD206 + macrophages and MFI of CD206 (G), and the percentage of IL-10 + macrophages and MFI of IL-10 (H). Data from (A) are means ± SEM for n = 5 from three independent experiments. Data from (C) and (D) are means ± SEM for n = 5 for the PBS group, n = 6 for the S3QEL 1.2 group, n = 7 for the CpG group, and n = 6 for the S3QEL 1.2 + CpG group. Statistical significance for survival analysis was determined by Mantel-Cox test. Statistical significance for tumor growth analysis was determined by two-way ANOVA; asterisks are for P < 0.05 in the group’s comparison. Data from (E) to (H) are means ± SD for n = 9 from one independent in vivo experiment. P values were calculated using two-tailed unpaired Student’s t test. Differences were statistically significant at * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: For in vivo samples, a Quantikine ELISA kit for mouse IL-10 (R&D Systems) was used according to the manufacturer’s instructions with appropriately diluted serum added to each plate.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, In Vivo, Flow Cytometry, Expressing, Fluorescence, Comparison, Two Tailed Test

Journal: Applied Sciences

Article Title: Development of Automated Exosome Isolation Method Using Epigallocatechin Gallate-Modified Magnetic Beads: Standardized Protocols for Diverse Biofluids

doi: 10.3390/app15116170

Figure Lengend Snippet: Figure 9. Comparative analysis of CD63 expression in exosomes isolated from various types of biological samples using Western blot and ELISA: (A) urine, (B) plasma, (C) serum, and (D) saliva.

Article Snippet: For urine and saliva samples, a human CD63 ELISA kit (Cusabio, Wuhan, China) was used following the manufacturer’s instructions.

Techniques: Expressing, Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Clinical Proteomics