Figure 4

Distribution of LNP-srRNA in mice after intravenous injection and in vitro expression of IL-12p70 by mouse tumor and primary cells after transfection with IL-12 srRNA. (A) Female C57BL6 mice were subcutaneously implanted with MC38 cells. 6 days later, Cy5-labeled LNP-srRNA-IL-12 was injected intravenously. The biodistribution of LNP-srRNA in selected organs and tumor was monitored at indicated timepoints with IVIS® Lumina III In Vivo Imaging System (n = 3). Scale bar = 10 mm. (B) Murine tumor cells (B16F10, MC38 and EMT6) and mouse primary cells (colon epithelial cells, and splenocytes) were used in this assay. 2 × 106 splenocytes or 8 × 104 other types of cells in 24 well plates were transfected with 50 or 500 ng srRNA encoding mouse IL-12p70 using Lipofectamine MessengerMax or LNP encapsulation (JCXH-211m). Supernatants were collected at 48 h post transfection for IL-12p70 determination by ELISA. Data were presented as mean ± SD, each dot represents one individual sample (n = 3).