Mediators of Inflammation / 2019 / Article / Fig 4

Research Article

Pregnancy-Associated Plasma Protein A Induces Inflammatory Cytokine Expression by Activating IGF-I/PI3K/Akt Pathways

Figure 4

Upregulation of MCP-1, TNF-α, and IL-6 expressions and productions by PAPP-A is dependent on the IGF-I/PI3-K/Akt signaling pathway in RAW264.7 macrophages. Cells were incubated for 24 h in the presence (+) or absence (−) of PAPP-A (200 ng/mL) with (+) and without (−) LY294002 (10 μM). In addition, RAW264.7 macrophages were transfected with control or IGF-IR siRNA and then incubated with PAPP-A (200 ng/mL) for 24 h. Total RNA was extracted, and RT-qPCR was performed to determine the expression of MCP-1, TNF-α, and IL-6 mRNA (A), and Western immunoblotting assays were conducted for protein expression of MCP-1, TNF-α, IL-6, and β-actin (b, c), and then ELISA were used to estimate MCP-1, TNF-α, and IL-6 concentrations in the culture medium (d, e, f). Similar results were obtained in three independent experiments. Data are the . vs. baseline.
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