Figure 6.

lnc-CCNL1-3:1 interaction with FOXO1 by enhancing its nuclear location

(A) Nuclear and cytoplasmic fractions of Enter- or CCNL-transfected cells were subjected to western blotting analysis to measure relative protein expression. The cytoplasmic tubulin and nuclear lamin A/C were used as controls. (B) Nuclear and cytoplasmic fractions of granulosa cells were subjected to quantitative real-time PCR to detect the intracellular localization of CCNL. The cytoplasmic ACTB and nuclear U6 were used as controls. (C) The efficiency of RNA immunoprecipitation (RIP) is shown as quantitative real-time PCR of U1 snRNA. (D) Enrichment of CCNL in granulosa cells was measured by quantitative real-time PCR from RIP with FOXO1 antibody or immunoglobulin G (IgG). IgG immunoprecipitation (IP) was used as a negative control. (E) Cells were incubated with CCNL (pCCNL) or Enter for 72 h. Subcellular localization of FOXO1 was visualized using anti-FOXO1 (green), and the nuclei were counterstained with DAPI (blue). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 correspond to two-tailed Student’s tests.