Fig. 6.

LEENE RNA promotes RNA Pol II binding and eNOS transcription. a–c, e HUVECs were treated with ATV for 24 h. The binding of RNA Pol II, KLF4, and MED1 to LEENE RNA was determined by RIP followed by qPCR (a–c). d Predicted secondary structure of LEENE RNA based on minimum free energy (MFE) and fragments complementary to ChIRP probes are labeled with numbers 1–10. Color scale shows the probabilities for every nucleotide to hold the structural position. Following ChIRP, interactions between LEENE RNA and respective DNA regions of LEENE and eNOS were detected by qPCR (in e). f, g Static ECs were transfected with scramble or LEENE LNA. The binding of RNA Pol II to eNOS promoter was determined by ChIP-qPCR with three primer sets flanking three regions upstream of eNOS TSS (in f). g Nascent RNA was captured in static ECs transfected with scramble or LEENE LNA. eNOS mRNA level was detected by qPCR. Data are presented as mean ± SEM, n = 5 in each group. *p < 0.05 compared with respective control in each experiment. In a–c, * denotes p < 0.05 compared with DMSO; in e, * indicates p < 0.05 between Ctrl and ATV using even-numbered probes; † denotes p < 0.05 between Ctrl and ATV using odd-numbered probes; in f and g, * means p < 0.05 between scramble vs. LNA groups. Student’s t test was applied