Figure 2: Atherosclerosis-related cell functions in circANRIL-overexpressing cells.

(a) Schematic of the vector construct for circANRIL stable and transient overexpression. RNA polymerase II (RNAP II). (b) ANRIL isoform expression in HEK-293 cells that stably express circANRIL, linANRIL or empty vector (three biological replicates each). PCR of reverse-transcribed RNA, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), house-keeping gene; OdT, oligo(d)T primers; RH, random hexamer primers. (c) Apoptosis and (d) proliferation in HEK-293 cell line stably overexpressing circANRIL (pool of 3 biological replicates, and 8 and 12 technical replicates per condition, respectively), compared with overexpression of an unrelated circular RNA (circHPRT1). Apoptosis (e) and proliferation (f) after RNAi against circANRIL or non-circANRIL or scrambled (SCR) siRNA control. Quadruplicate measurements per condition. *P<0.05; **P<0.01; ***P<0.001. Comparison of multiple groups was done using analysis of variance, and Tukey was performed as post hoc test in c,e. Two-tailed unpaired Student’s t-test in d,f. Data are given as mean±s.e.m.