USP30-AS1 binds to HnRNPF and HnRNPF regulates breast cancer cell proliferation. (IMAGE)
Caption
(A) FISH assay was conducted to determine the localization of USP30-AS1 in MDA-MB-231 cells. The representative images were acquired using confocal microscopy.
(B) Nuclear-cytoplasmic fractionation was performed in MDA-MB-231 and MCF-7 cells. The expression of USP30-AS1 in the nucleus and cytoplasm was evaluated by qRT-PCR. Neat1 and MT-CYB were used as markers for the nuclear and cytoplasmic localized genes, respectively.
(C) The predicted binding scores of USP30-AS1 with HnRNPF, RPS2L, RPL19, and HnRNP-U proteins based on mass spectrometry results. The results of RNA pull-down assay in MDA-MB-231 cells confirmed the interaction between USP30-AS1 and HnRNPF.
(D) Confocal images showed the colocalization of USP30-AS1(red) and HnRNPF (green) in MDA-MB-231 cells.
(E) RIP assay showed the interaction between HnRNPF and USP30-AS1 in MDA-MB-231 cells.
(F) The effect of HnRNPF on breast cancer cell proliferation was conducted in MDA-MB-231 cells transfected with HnRNPF siRNA or HnRNPF plasmid.
(G) The impact of HnRNPF on the proliferation capacity of MDA-MB-231 cells was evaluated by colony formation assays. (H) Flow cytometry analysis was conducted to determine the impact of HnRNPF on cell cycle progression in MDA-MB-231 cells.
(I) EdU assays were performed in MDA-MB-231 cells with HnRNPF knockdown or
(J) overexpression, and the percentage of EdU positive cells was quantified based on EdU staining.
Credit
Yapei Jiang, Weijie Liao, Qilei Xin, Ruonan Wang, Guanglan Lin, Jia Li, Zijian Yang, Shiyue Yang, Haowei Zhang, Xiaolin Li, Qian Peng, Yaou Zhang, Weidong Xie, Naihan Xu
Usage Restrictions
Credit must be given to the creator. Only noncommercial uses of the work are permitted. No derivatives or adaptations of the work are permitted.
License
CC BY-NC-ND