TNF-α induced by TSP50 is crucial for affecting macrophage activities.

<p>(A-B) TSP50-o/e cells were transfected with the indicated shRNA plasmids. After 24h, the mRNA level of TNF-α (A) or cytokines (B) in these cells were analyzed by real-time PCR. (C) dTHP-1 cells or mouse peritoneal macrophages were cultured with CM from TNF-α knock-down TSP50-o/e cells or control cells for 24h. The mRNA expression of cytokines and macrophage phenotypic markers were determined by real-time PCR. (D) Mouse peritoneal macrophages were cultured with CM from TNF-α knock-down TSP50-o/e cells or control cells for 24h. CM from macrophages was collected and subjected to ELISA to detect the secretion of cytokines. (E) Mouse macrophages were cultured with CM from TNF-α knock-down TSP50-o/e cells or control cells for 24h. The macrophages were collected and lysed, and the protein level of macrophage phenotype markers were analyzed by western blotting. (F) Phagocytosis capacities of dTHP-1 cells (up) or mouse peritoneal macrophages (down) were measured after treatment with CM from TNF-α knock-down TSP50-o/e cells or control cells. GAPDH was used as the internal control to check the efficiency of cDNA synthesis and PCR amplification. Data are shown as mean ± SD of three replicates in three independent experiments.* p<0.05, **p<0.01.