Figure 1

Detection of hsa-miR-29a, b, c in different cell types. (A) Schematic representation of the HIV-1 genome. Potential target sites complementary to hsa-miR-29a and b are marked with an arrow. (B) Schematic representation of the method of detection and the oligonucleotides used to capture the miRNA(s). The oligonucleotides (blue) prime sequence specific extension (green) of each miRNA due to differences at the 3' end of the oligonucleotide-miRNA hybrid. The extension product is radiolabeled by the introduction of alpha-P³²-dCTP into the product at the positions underlined. The T tail of varying lengths at the 5' end is used to improve the resolution of the products. (C) Expression of hsa-miR-29a, b, c in PBMC (upper panel), HeLa (middle panel) and HEK293T (lower panel) cells. HEK293T cells show much lower expression level of hsa-miR-29a and b than PBMCs or HeLa cells. (D) U6 RNA was detected using a similar approach to establish equal input of RNA. Product sizes (nucleotides, nt) including length of T tails are indicated on the right. M: radiolabeled marker.