Journal: Frontiers in Immunology

Article Title: Pan-cancer analysis identifies tRNA modification enzyme CTU2 as a novel tumor biomarker and its role in immune microenvironment

doi: 10.3389/fimmu.2025.1547794

Figure Lengend Snippet: CTU2 knockdown impairs LIHC and KIRC progression in vivo and in vitro . (A) Western blot confirming CTU2 knockdown in huh-7 and 786-O. Control (untransfected wild-type cells), NC (lentiviral empty vector group), Sh-CTU2 (lentivirus-mediated CTU2 knockdown). The lower graph shows the grey value of CTU2 protein levels, normalized to the corresponding GAPDH levels. The experiment was independently repeated three times (** P < 0.01); (B) Colony-formation assay of CTU2 knockdown and control huh-7 and 786-O, representative images (left panel), and the quantitative analysis (right panel, *** P < 0.001); (C) EdU proliferation assay (upper) and the quantitative analysis (lower) of CTU2 knockdown and control cells (* P <0.05, ** P < 0.01); (D) AnnexinV/PI apoptosis assay (upper) and the quantitative analysis (lower) of CTU2 knockdown and control cells (* P <0.05, ** P < 0.01); (E) Matrigel invasion assay of CTU2 knockdown and control 786-O (upper) and huh-7 (lower) cell, representative images (left panel), and quantification analysis (right panel, ** P < 0.01, *** P < 0.001, **** P < 0.0001); (F) Transwell cell migration analysis of CTU2 knockdown and control 786-O (upper) and huh-7 (lower) cell, representative images (left panel), and quantification analysis (right panel); (G) Representative picture of tumors in xenograft nude mice model subcutaneously implanted with CTU2 knockdown and control huh-7 cells; (H) Xenograft tumor weigh (left, n = 5, ** P < 0.01) and xenograft tumor growth curve (right, n = 5, * P < 0.05).

Article Snippet: For apoptosis assays, the Alexa Fluor 488 Annexin V/PI Cell Apoptosis Kit (Vazyme, A211-01, China) was used according to the manufacturer’s instructions.

Techniques: Knockdown, In Vivo, In Vitro, Western Blot, Control, Plasmid Preparation, Colony Assay, Proliferation Assay, Apoptosis Assay, Invasion Assay, Migration