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

Journal: Translational Oncology

Article Title: Ferroptosis induction enhances anti-PD-1 efficacy in NSCLC via HIF-1α/PD-L1 modulation

doi: 10.1016/j.tranon.2026.102685

Figure Lengend Snippet: Ferroptosis inducers cause cell death and affect PD-L1 expression, which can be inhibited by iron chelators. (A, B) A549 and H1299 cells were treated with erastin or RSL3 at different concentration gradients for 24 h, and cell viability was detected using the CCK-8 kit. Subsequently, cells were co-treated with DFO (100 μM), ferrostatin-1 (2 μM), and the aforementioned ferroptosis inducers at different concentrations for another 24 h, with cell viability re-detected by CCK-8.(C) A549 and H1299 cells were treated with erastin (20 μM) and RSL3 (5 μM), respectively, for 0, 3, 6, 12, 18, and 24 h. PD-L1 expression was measured by qPCR.(D) After A549 and H1299 cells were treated with erastin (20 μM) for 24 h, they were further treated with DFO (100 μM) and ferrostatin-1 (2 μM) alone or in combination. PD-L1 expression in cells was then detected. (E) A549 and H1299 cells were treated with erastin (20 μM) for 24 h, and intracellular ROS was detected by flow cytometry. CCK-8, cell counting kit-8; DFO, deferoxamine; qPCR, quantitative polymerase chain reaction; PD-L1, programmed death-ligand 1; ROS, reactive oxygen species. *** p < 0.001.

Article Snippet: Reagents and Antibodies Cell culture media and reagents: Ham's F-12 K, DMEM, RPMI-1640 (Servicebio), BCA protein assay kit, CCK-8 kit, RIPA lysis buffer (Servicebio), Lovastatin, Fluvastatin, RSL3, Erastin (MCE), deferoxamine mesylate, and Ferrostatin-1 (MCE).

Techniques: Expressing, Concentration Assay, CCK-8 Assay, Flow Cytometry, Cell Counting, Real-time Polymerase Chain Reaction