Fig 1: Increased platelet-mediated promotion of C5 transcription requires PSGL-1. (A-B) GO-BP analysis (cell adhesion) of the genes in TAMs isolated from mice treated with or without TPO. (C) KEGG analysis (cell adhesion) of the genes in TAMs isolated from mice treated with or without TPO. (D) Real-time RT-PCR was used to quantify the levels of Psgl1, Itgb1, Itgb4, Itgb5, Itga5, and Ptafr mRNA (n = 3). The data shown are representative of a single experiment. (E) BMDMs were treated with or without platelets for 6 h. Then, cell lysates were harvested and subjected to immunoblotting. (F) Tumor volumes and tumor weights were examined (n = 7). (G) The serum concentration of C5a was measured by ELISA (n = 7). (F-G) The data shown are representative of a single experiment. (H-I) The relative mRNA levels of C5, Il10, and Arg1 were examined in BMDMs treated without or with platelets (H) or in the absence/presence of CD62P (I) for the indicated times. The data shown are representative of one of two independent experiments. (J-K) BMDMs were transfected with a negative control siRNA or an siRNA against PSGL-1. After 24 h, cell lysates were harvested and subjected to immunoblotting to examine the indicated proteins (K) and real-time RT-PCR to quantify the Psgl1 and C5 mRNA levels. The cell culture medium was harvested, and C5a was examined. (J-K) The data shown are representative of one of three independent experiments. **P < 0.01; *P < 0.05; n.s., not significant.