Figure 3.

Cytokine expression was altered in the RAW264.7 cells by transfection of the target genes (gra15_II or rop16_I/III). The culture supernatants were collected and analyzed for TNF-α (a), NO (b), IL-10 (c), urea (d), IL-6 (e) and TGF-β1 (f) production by the transfected RAW264.7 and control RAW264.7 cells. The TNF-α and NO levels were enhanced and IL-6 was decreased in the GRA15_II-RAW264.7 cells, whereas the IL-10, urea, IL-6 and TGF-β1 levels were increased in the ROP16_I/III-RAW264.7 control cells compared with the LV-RAW264.7 cells. The transfected macrophage-related proteins Flag, iNOS, Arg-1 and MMP13 were detected by western blotting (g). The Flag tag was expressed with an approximate molecular weight of 58 kD (GRA15_II-RAW264.7 cells) and 80 kD (ROP16_I/III-RAW264.7 cells). Higher iNOS and MMP13 expression levels were detected in the GRA15_II-RAW264.7 cells compared with the LV-RAW264.7 cells. Correspondingly, escalated Arg-1 expression was detected in the ROP16_I/III-RAW264.7 cells compared to the LV-RAW264.7 cells (**P<0.01, ***P<0.001, n=4). Arg-1, arginase-1; IL, interleukin; iNOS, inducible nitric synthase; MMP13, matrix metalloproteinase 13; NO, nitric oxide; TNF, tumor necrosis factor; TGF-β1, transforming growth factor beta 1.