Skip to main content
. 2012 Feb 29;287(17):13899–13910. doi: 10.1074/jbc.M111.301275

FIGURE 9.

FIGURE 9.

MiR-219–5p regulation on PGR via lncRNA-PGR-3p. A, sequences of predicted miR-219–5p binding site in the lncRNA-PGR-3p non-coding RNA. B, target validation by luciferase assay in rhesus monkey and human. The 3′-UTR fragments derived from rhesus monkey and human were cloned and subjected to luciferase assay in rhesus LLC-MK2 cells and human ECC-1 cells, respectively. C, point mutation analysis of miR-219–5p seed binding sequence. The seed binding site (ACAATCA) was mutated into AAACTAC. Cells were co-transfected with miR-219–5p precursor. D, efficiency of lncRNA-PGR-3p RNAi on lncRNA-PGR-3p RNA level determined by qRT-PCR. The siRNA sequence designed for rhesus lncRNA-PGR-3p was 5′-GCUGUAAGUUUGAGUUGAUdTdT-3′. E, Western blot analysis of PGR-A protein expression after rhesus LLC-MK2 cells were treated with lncRNA-PGR-3p siRNA. F, level of endogenous PGR-A protein measured by Western blot after rhesus LLC-MK2 cells were transfected with the anti-miR-219-5p inhibitor or control. G, indirect regulation of PGR by miR-219-5p via lncRNA-PGR-3p. When rhesus LLC-MK2 cells were co-transfected with lncRNA-PGR-3p siRNA, miR-219–5p down-regulation on PGR luciferase activity was abrogated.