factor 1α quantikine elisa kit  (R&D Systems)

Journal: International Journal of Oncology

Article Title: Cancer-associated fibroblast-induced M2-polarized macrophages promote hepatocellular carcinoma progression via the plasminogen activator inhibitor-1 pathway

doi: 10.3892/ijo.2021.5239

Figure Lengend Snippet: PAI-1 is the key factor secreted by TAMs following CAF-CM stimulation and promotes tumor malignant behavior in vitro . (A) Original image and (B) spot pixel value from a cytokine array revealed the profiles of paracrine factors in the M0-CM, TAM (Ca)-CM and TAM(CAF)-CM. (C) Western blot and (D) ELISA results indicated that PAI-1 was upregulated in the TAM(CAF) group compared with that in the M0 and TAM(Ca) groups. (E) Cell Counting Kit-8 and (F) colony formation assays showed that the inhibition of PAI-1 in TAM(CAF)-CM suppressed the enhanced proliferation of Huh-7 cells. (G) Wound healing (scale bar, 400 µ m) and (H) Transwell and (I) Matrigel (scale bar, 200 µ m) assays indicated that the inhibition of PAI-1 in the TAM(CAF)-CM group suppressed the enhanced migration and invasion of the Huh-7 cells. * P<0.05; ** P<0.01. CAF, cancer-associated fibroblast; CM, conditioned medium; CXCL12, C-X-C motif chemokine ligand 12; CXCL5, C-X-C motif chemokine ligand 5; PAI-1, plasminogen activator inhibitor-1; PDGF-AA, platelet derived growth factor-AA; TAM, tumor-associated macrophage; TIM-3, T-cell immunoglobulin mucin 3; M0, macrophages; NS, not significant.

Article Snippet: The Human IL-6 Quantikine ELISA kit (cat. no. D6050), Human Serpin E1/PAI-1 Quantikine ELISA kit (cat. no. DSE100) and Human CXCL12/stromal cell-derived factor 1α Quantikine ELISA kit (cat. no. DSA00) were purchased from R&D Systems Inc., to detect the concentrations of IL-6, PAI-1 and CXCL12 in the CM, according to the manufacturer's instructions.

Techniques: In Vitro, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Counting, Inhibition, Migration, Derivative Assay

Journal: International Journal of Oncology

Article Title: Cancer-associated fibroblast-induced M2-polarized macrophages promote hepatocellular carcinoma progression via the plasminogen activator inhibitor-1 pathway

doi: 10.3892/ijo.2021.5239

Figure Lengend Snippet: CAF-derived CXCL12 induces the secretion of PAI-1 in TAM(CAF). (A) Original image and (B) spot pixel value from a cytokine array to identify the different patterns of molecules in cancer-CM and CAFs-CM. CXCL12 gene expression and its secretion were increased in CAFs compared with that in the Huh-7 and Lx-2 cells following (C) RT-qPCR and (D) ELISA. (E) CXCR4 gene expression in M0, TAM(Ca) and TAM(CAF) was analyzed using RT-qPCR. (F) RT-qPCR and (G) ELISA results demonstrated that the gene expression level and secretion of PAI-1 were decreased in TAM(CAF) after CXCL12 neutralization in CAF-CM, respectively. (H) Schematic representation of the proposed interactions between HCC, TAMs and CAFs. * P<0.05; ** P<0.01. CAF, cancer-associated fibroblast; CM, conditioned medium; CCL5, C-C motif chemokine ligand 5; CXCL12, C-X-C motif chemokine ligand 12; CXCR4, C-X-C Motif chemokine receptor 4; HCC, hepatocellular carcinoma; HSCs, hepatic stellate cells; PAI-1, plasminogen activator inhibitor-1; PDGF-AA, platelet derived growth factor-AA; RT-qPCR, reverse transcription-quantitative PCR; TAM, tumor-associated macrophage; Ab, antibody.

Article Snippet: The Human IL-6 Quantikine ELISA kit (cat. no. D6050), Human Serpin E1/PAI-1 Quantikine ELISA kit (cat. no. DSE100) and Human CXCL12/stromal cell-derived factor 1α Quantikine ELISA kit (cat. no. DSA00) were purchased from R&D Systems Inc., to detect the concentrations of IL-6, PAI-1 and CXCL12 in the CM, according to the manufacturer's instructions.

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Neutralization, Reverse Transcription, Real-time Polymerase Chain Reaction