Journal: Genes & Diseases
Article Title: Nuclear and cytoplasmic USP30-AS1 coordinately regulate breast cancer progression through HnRNPF/p21 and EZH2/c-Myc/p21 axes
doi: 10.1016/j.gendis.2025.101684
Figure Lengend Snippet: USP30-AS1 knockdown inhibits breast cancer cell proliferation both in vitro and in vivo . (A) The knockdown efficacy of USP30-AS1 siRNA and shRNA was validated by qRT-PCR analysis. (B) The effect of USP30-AS1 knockdown on MDA-MB-231 cell proliferation was evaluated by CCK-8 assays. (C) The impact of USP30-AS1 knockdown on the proliferative capacity of MDA-MB-231 cells was assessed by colony formation assays. (D) Flow cytometry analysis was performed to determine the effect of USP30-AS1 knockdown on the cell cycle progression of MDA-MB-231 cells. (E) EdU assays were conducted in MDA-MB-231 cells with USP30-AS1 knockdown. (F) A xenograft tumor experiment was conducted using three MDA-MB-231 cell lines: shNC, shUSP30-AS1#1, and shUSP30-AS1#2. (G) Tumor growth and tumor weight were assessed every two days. (H) The actual photo of tumors for each group was collected at the end of the experiment. (I) Hematoxylin and eosin staining of tumor tissues from nude mice. (J) Immunohistochemistry assays were performed to determine the impact of USP30-AS1 knockdown on Ki67 expression within xenograft tumor tissues.
Article Snippet: The cells were seeded into a 96-well plate (2 × 10 3 /well) and cultured for 0, 24, 48, 72, and 96 h. Cells were treated with 10% CCK-8 solution (MedChemExpress, # HY-K0301) and incubated for 2 h. OD 450 was determined using a microplate reader.
Techniques: Knockdown, In Vitro, In Vivo, shRNA, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Staining, Immunohistochemistry, Expressing