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. 2014 May 15;42(12):7981–7996. doi: 10.1093/nar/gku424

Figure 1.

Figure 1.

Absolute quantification of mature miRNAs from KSHV in various cell lines. (A) To quantify the absolute amount of each miRNA in the different cell types, a standard RNA oligonucleotide, identical to the tested miRNA, was loaded on the same gel at increasing amounts (0.1–10 fmol) along with 5 μg (DG-75-EGFP, DG-75-K10/12 and BCBL-1) or 2.5 μg (BC-3) of total RNA. Representative northern blots for each of the miRNA tested (n = 3–4) are shown. As a negative control, total RNA from a non-infected cell-line DG-75-EGFP was used. miR-16 was probed as a loading control. (B) pcDNA5 expression plasmids with the entire miRNA cluster (cluster) or only single pre-miRNA (unique) were transfected in HEK293Grip cells and total RNA analyzed by northern blot for the indicated miRNAs. miR-16 was probed as a loading control, and the KSHV miRNA signals quantified and normalized relative to the cluster-transfected cells, which was set to 1.