Figure 2.
In vivo visualization of expression of pri-miR-23a transcripts and activity of mature forms of miR-23a in nude mice. (A) HeLa cells, (B) 293 cells, and (C) undifferentiated P19 cells were transfected with miR23P639/Fluc and CMV/Gluc/3xPT_mir23, and were grafted (1 Х 107 cells) to right side of mice. The same cells transfected with pGL3_basic and CMV/Gluc were grafted to left side as controls. Quantitative data from imaging in vivo were obtained by ROI analysis and are shown next to each small-animal picture. Gluc images were acquired first with coelenterazine, and then Fluc images were then acquired with D-luciferin. When viewing Fluc images, pay attention to increased bioluminescence on right side of grafted cells expressing pri-miR-23a transcripts. When viewing Gluc images, note the decreased bioluminescence on right side compared with luciferase activity on left-sided CMV/Gluc controls, disclosing mature miR-23a activities. Fluc activities increased in all animals on right side, and Gluc activities decreased in HeLa and P19 cells, but decreased the least in 293 cells transfected with CMV/Gluc/3xPT_mir23 on right side.