656158.fig.002a
(a)
656158.fig.002b
(b)
Figure 2: (a) Accumulation of FAM-labeled artificial box C/D RNAs in human cells. MCF-7 cells transfected with FAM-labeled RNA8 analogue using Lipofectamine were analyzed with flow cytometry. “P2” population represents MCF-7 cells positively stained with FAM-labeled RNA, with “P1” population representing nonstained cells. (b) Presence of radiolabeled artificial box C/D RNA in total RNA from transfected MCF-7 cells. Radiolabeled RNA8 was transfected into cells using Lipofectamine, and the cells were cultured for time intervals from 3 h to 72 h (lanes 1–5). Total cellular RNA was isolated and analyzed in a 10% denaturing polyacrylamide gel followed by autoradiography of the radiolabeled RNA fragments. The RNA loading was normalized by spectrometry, and 3 µg of total cellular RNA was used per lane. Lane 6 intact radiolabeled artificial box C/D RNA. The arrows indicate marker mobility (in parallel sequencing reaction product were loaded on the same gel).