Figure 2
EVs promote TGFβ production in macrophages in vivo. (a) mRNA level of TGFβ expressed by macrophages in vivo: EVs (n = 4) or PBS (n = 4) were intraperitoneally injected into C57BL/6 mice, and peritoneal macrophages were harvested at 24 hr and quantified TGFβ and GAPDH mRNA. Data of two independent experiments were combined. (b) Protein level of TGFβ produced by macrophages in vivo: EVs (n = 3) or PBS (n = 3) were intraperitoneally injected to wild type C57BL/6 mice, and peritoneal macrophages were harvested at 24 hr and further cultured in serum-free medium for next 24 hr. The TGFβ in supernatant was quantified by ELISA. Data were representative for three independent experiments. (c) TGFβ production in macrophages challenged with LPS: EVs (n = 3) or PBS (n = 3) were intraperitoneally injected into C57BL/6 mice, and then challenged intraperitoneally with LPS (200 μg per mouse) 4 hr later. Peritoneal macrophages were harvested at 16 hr and further cultured in serum-free medium for next 24 hr. TGFβ in supernatant were quantified by ELISA. Data were representative for two independent experiments. (d) Plasma levels of TNFα in mice challenged with LPS: EVs (n = 3) or PBS (n = 3) was intraperitoneally injected into C57BL/6 mice, and mice were challenged intraperitoneally with LPS 4 hr later. Plasma was harvested at 16 hr and quantified for TNFα by ELISA. Data were representative for two independent experiments. (e) TNFα production in macrophages challenged with LPS: EVs (n = 3) or PBS (n = 3) were intraperitoneally injected into C57BL/6 mice, then challenged with intraperitoneal injection of LPS 4 hr later. The peritoneal macrophages were harvested at 16 hr and restimulated with LPS in serum-free medium for next 24 hr. The level of TNFα in supernatant was quantified by ELISA. Data were representative for two independent experiments.
