Figure 3
From: In vitro and in vivo targeted delivery of IL-10 interfering RNA by JC virus-like particles
Suppression of IL-10 expression in RAW 264.7 cells by two IL-10 shRNA preparations. Cells were cultured with either IL-10 shRNA (IL-10i-1 or IL-10i-2) or irrelevant shRNA for 24 h. After washing, gene expression was analyzed by real-time PCR. Represented are the mRNA levels of target gene relative to those of 18 s rRNA. (A) Suppression of IL-10 expression was analyzed by real-time PCR. (B) IL-10 shRNA did not cross-react with TNF-α mRNA and influence the expression of TNF-α in RAW 264.7 cells. Relative TNF-α expression was analyzed by Real-time PCR. (C) Suppression of IL-10 expression by various doses of IL-10 shRNA in RAW 264.7 cells. (D) RAW 264.7 cells (2 × 105) were pre-treated with LPS (2 μg/ml) for 90 min. Cells were then cultured with either IL-10 shRNA (IL-10i-2) or irrelevant shRNA for 24 h. After washing, IL-10 expression was analyzed by real-time PCR. Represented are the mRNA levels of IL-10 relative to those of 18 s rRNA. *: P < 0.05; ***: P < 0.005. (E) RAW 264.7 cells (2 × 105) were pre-treated with LPS (2 μg/ml) for 90 min. Cells were then cultured with either IL-10 shRNA (IL-10i-2) or irrelevant shRNA for 36 h. After 36 h, conditioned medium was collected for analysis of TNF-α expression by ELISA. (F) RAW 264.7 cells (2 × 105) were pre-treated with LPS (2 μg/ml) for 90 min. Cells were then cultured with either IL-10 shRNA (IL-10i-2) or irrelevant shRNA for 0, 6, 12, 24, 36, 48 and 60 hours. After 0, 6, 12, 24, 36, 48 and 60 hours, conditioned medium was collected for analysis of IL-10 expression by ELISA. *: P < 0.05.