Figure 2.

LncRNA-RNCR3 is upregulated in aortic atherosclerotic lesion and its knockdown aggravates atherosclerosis in vivo. (a) RNCR3 expression in the aorta of 5-month-old male ApoE^−/− and C57B/6J mice was determined by qRT-PCRs and normalized to the expression of GAPDH. The data were expressed as relative mRNA expression compared with average expression in wild-type group (WT). WT, wild type C57B/6J mice. E/L-lesion and E/L-normal: aorta segments with atherosclerotic lesions or without lesion (normal) from apoE^−/− mice, respectively (*P<0.05). (b) RNCR3 expression was detected in atherosclerotic lesions and non-lesional aortic intimal tissues from human aortas (*P<0.05). (c–f) Four-week-old male ApoE^−/− mice were fed with high-fat diet containing 0.15% cholesterol and 20% fat for 16 weeks. They were received a subcutaneous injection of scrambled shRNA (Scr) or RNCR3 shRNA viral vector (R), or PBS. shRNA injection was started at 4 weeks after feeding with high-fat diet. Viral vector was injected once every 2 weeks. Representative en face Oil red O staining in the aortas of PBS-, scrambled shRNA-, and RNCR3 shRNA-injected mice. Scale bar, 0.5 cm. Atherosclerotic lesions quantification in en face aortas was expressed as the percentage of lesions relative to total aortic area (C and D, n=6 per group). Representative oil red O staining of aortic sinus in PBS-, scrambled shRNA-, and RNCR3 shRNA-injected mice. Scale bar, 300 μm. Aortic sinus lesion quantification was shown as the change compared with PBS-injected group. *P<0.05 versus PBS-injected group (e and f, n=6 per group)