Fig. 2

LncRNA Oasl2-209 directly binds to miR-23a/b and regulates miR-23a/b expression. a L929 cells were treated with or without HT-DNA (5 µg ml⁻¹) for 6 h, and total RNA was then extracted and subjected to high-throughput sequencing. Heat map showing the differentially enriched lncRNAs with adjusted p values of less than 0.05 (padj < 0.05). b qPCR validation of eleven elevated lncRNAs in L929 cells treated as described in a. c Schematic description of the predicted interactions between the binding site of Oasl2-209 and miR-23a/b. The binding site sequence is marked in red, and the predicted minimum free energy values of the hybridizations are shown. L929 cells were infected with HSV-1 (MOI = 1) (d) or stimulated with HT-DNA (5 µg ml⁻¹) (e) for the indicated times. Then, the level of Oasl2-209 was measured by qPCR. f Fragments of Oasl2-209 harboring wild-type (Oasl2-209-WT) or mutant (Oasl2-209-MUT) miR-23a/b target sequences were cloned into pmirGLO reporter plasmids. The mutated nucleotides are marked in bold. g HEK293 cells were transfected with miR-23a/b mimics or control mimics along with the indicated reporter plasmids for 24 h. Then, the relative luciferase activities were analyzed. MEFs (h) and L929 cells (i) were transfected with biotinylated miR-23a/b probes for 48 h and were then harvested for a biotin-avidin pulldown assay. RNA was extracted, and the level of Oasl2-209 was measured by qPCR. j L929 cells were harvested and incubated with Oasl2-209 probe-coated streptavidin beads for a pulldown assay. RNA was extracted, and the levels of miR-23a/b were measured by qPCR. k L929 cells were transfected with the indicated plasmids, and the levels of Oasl2-209 and miR-23a/b were measured by qPCR. l L929 cells transfected with the indicated siRNAs were stimulated with HT-DNA for 6 h, and the levels of Oasl2-209 and miR-23a/b were then measured by qPCR. The data are representative of three independent experiments (mean ± SD). *p < 0.05, **p < 0.01