Mediators of Inflammation / 2019 / Article / Fig 3

Research Article

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

Figure 3

PAPP-A induces the IGF-I/PI3-K/Akt pathway in macrophages. (a, c, e) RAW264.7 macrophages were treated with various concentrations of PAPP-A for 24 h, respectively. (b, d, f) Cells were treated with 5 mg/mL BSA for 24 h or with 200 ng/mL PAPP-A for different durations, respectively. (a, b) IGF-I, IGF-IR, and IGFBP-5 mRNA expressions were measured by RT-qPCR. (c, d, e, f) IGF-I, IGF-IR, and IGFBP-5 protein expressions were measured by Western immunoblotting assays. (g, h) Free IGF-I concentrations in the culture medium was estimated with ELISA. (i, j) RAW264.7 macrophages were transfected with control or IGF-IR siRNA and then incubated with PAPP-A (200 ng/mL) for 24 h. (i) Protein samples were immunoblotted with anti-IGF-IR or anti-β-actin antibodies. (j) Western analysis for phosphorylated Akt and PI3-K p85 subunit. Similar results were obtained in three independent experiments. Data are the . vs. baseline, vs. baseline, and vs. baseline.

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