Fig. 2: CDK2/cyclin E1 controls the nuclear export of NF90 to stabilize cyclin E1 mRNA and promotes cell proliferation in HCC cells.
From: NF90 stabilizes cyclin E1 mRNA through phosphorylation of NF90-Ser382 by CDK2
a NF90-WT and NF90-S382A display different nuclear export abilities. GFP tagged NF90-WT or NF90-S382A were transfected into HeLa cells. After 24 h, cells were fixed and stained with DAPI. C, cytoplasm; N, nuclear. Columns, mean (n = 30); bar, s.d. **p < 0.01. b Detection of nuclear export of GFP-NF90 after transfection with CDK2/cyclin E1 or not. C, cytoplasm; N, nuclear. Columns, mean (n = 30); bar, s.d. ***p < 0.001. c Huh 7 cells stably expressing NF90-S382A, NF90-WT or the control were constructed. Cyclin E1 expression in the stable lines was detected by WB. d The Huh 7 cells stably expressing NF90-S382A was used to examine the half-life of cyclin E1 mRNA. Points, mean (n = 3); bar, s.d. **p < 0.01. e Increasing roscovitine significantly decreased cyclin E1 mRNA half-time in Huh 7 cells stably expressing NF90-WT instead of NF90-S382A after 1.5 h incubation with actinomycin D. Points, mean (n = 3); bar, s.d. **p < 0.01. f Cell cycle analysis was carried out in Huh7 cells stably transfected with vector, NF90-WT, or NF90-S382A. The fraction of viable cells in the different cell cycle phases was analyzed by flow cytometry. Columns, mean (n = 3); bar, s.d. **p < 0.01; ***p < 0.001. g Colony formation assays were performed to assess the growth ability of Huh7 cells transfected with vector, NF90-WT, or NF90-S382A. Left, representative images of colony formation assay; right, analysis of the colony numbers. Columns, mean (n = 3); bar, s.d. ***p < 0.001.
