Research Paper Volume 13, Issue 4 pp 5383—5402

CircMRE11A_013 binds to UBXN1 and integrates ATM activation enhancing lens epithelial cells senescence in age-related cataract

Knock-down of circMRE11A enhanced viability of SRA01/04 cell lines in vitro. (A). Designing three circMRE11A-specifific small interfering RNAs (siRNAs) targeted the back-splice junction sequence to knock down the expression levels of circMRE11A in LECs. Determining by qRT-PCR, the level of circMRE11A decreased after transfecting cells with siRNA-02 compared with the other siRNAs. *pB). SiRNA-02 targeting the back-splice junction of the circMRE11A which did not affect the expression of MRE11A mRNA. (C). CCK-8 showed that silencing of circMRE11A enhanced cell viability in the cell lines compared with controls. *pD). Flow cytometry demonstrated that silencing of circMRE11A promoted G1 entering S which means the progression of cells into the DNA synthesis phase (S phase). (E). Western blot showed that knockdown of circMRE11A deceased expression of ATM-S1981p, but not other proteins ATM, MRE11A, p53 and p21. Densities of bands were quantified by Image J software. Tubulin or GAPDH levels were measured in parallel, served as internal controls. *p

Figure 4. Knock-down of circMRE11A enhanced viability of SRA01/04 cell lines in vitro. (A). Designing three circMRE11A-specifific small interfering RNAs (siRNAs) targeted the back-splice junction sequence to knock down the expression levels of circMRE11A in LECs. Determining by qRT-PCR, the level of circMRE11A decreased after transfecting cells with siRNA-02 compared with the other siRNAs. *p<0.05. (B). SiRNA-02 targeting the back-splice junction of the circMRE11A which did not affect the expression of MRE11A mRNA. (C). CCK-8 showed that silencing of circMRE11A enhanced cell viability in the cell lines compared with controls. *p<0.05. (D). Flow cytometry demonstrated that silencing of circMRE11A promoted G1 entering S which means the progression of cells into the DNA synthesis phase (S phase). (E). Western blot showed that knockdown of circMRE11A deceased expression of ATM-S1981p, but not other proteins ATM, MRE11A, p53 and p21. Densities of bands were quantified by Image J software. Tubulin or GAPDH levels were measured in parallel, served as internal controls. *p<0.05.