Fig. 5

LINC01592 binds with E2F6 and accelerates the nuclear translocation of E2F6. A. The protein extracted from LINC01592 pulldown assays was analyzed by MS. B. According to RIP assays with anti-E2F6 antibodies, E2F6 interacted with LINC01592 in Eca-109/PECC. Top, agarose electrophoresis of PCR product. Bottom, qPCR result of RIP assay. C. The antisense sequences of LINC01592 served as negative controls for the WB detection of the protein obtained from LINC01592 pulldown assays. D. Representative images of E2F6 subcellular localization in Eca-109 under different treatments are shown in IF staining. Anti-E2F6 (red) and DAPI (blue) were used to stain all cells. Analysis of the average fluorescence intensity of E2F6 and DAPI signals was conducted using ImageJ. E. WB analysis of Eca-109 nuclear and cytosolic lysates was performed after knockdown or overexpression of LINC01592. F. Co-IP assays were performed on LINC01592 knockdown, LINC01592 overexpression, and control groups to determine the E2F6–NBR1 interaction. The means ± SDs are provided (n = 3). **P < 0.01 and ***P < 0.001 according to two-tailed Student t-tests.