Journal: Molecular Cancer

Article Title: CircZFR/YTHDF3 axis drives lymph node metastasis in cervical cancer via FASN translation

doi: 10.1186/s12943-025-02424-5

Figure Lengend Snippet: CircZFR Facilitates FASN Protein Translation via the YTHDF3-Dependent Mechanism. A Western blot analysis of FASN protein expression in HeLa and C33A cells treated with siYTHDF3#1 or siYTHDF3#2. B After treatment with MG132 for indicated times, protein levels of FASN were tested by Western blot analyses of HeLa cells treated with siCtrl, siYTHDF3#1, or siYTHDF3#2. C Coimmunoprecipitation by anti-eIF4A3 verified that the combination between YTHDF3 and eIF4A3 increased after overexpression circZFR and decreased after knocking down circZFR. D RIP assay using eIF4A3 antibody to test the combination between eIF4A3 protein and FASN mRNA in HeLa cells with or without circZFR overexpression. E MeRIP-qPCR assay validated the three m 6 A sites in FASN mRNA. F Schematic image of YTHDF3-WT plasmids, and YTHDF3-MUT plasmids containing m 6 A motif mutations in the CDS region. G Western blot assay tested the FASN protein expression in HeLa cells transfected with empty vector, YTHDF3-WT, or YTHDF3-MUT plasmids. WT, wild type; MUT, mutant. Data in A, B, C, D, E, and G are mean value ± SEM of three biological replicates, student t test, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Cell lysates were incubated with beads conjugated to YTHDF3 or eIF4A3 antibodies (Proteintech).

Techniques: Western Blot, Expressing, Over Expression, Transfection, Plasmid Preparation, Mutagenesis

Journal: Molecular Cancer

Article Title: CircZFR/YTHDF3 axis drives lymph node metastasis in cervical cancer via FASN translation

doi: 10.1186/s12943-025-02424-5

Figure Lengend Snippet: CircZFR Accelerates Lymph Node Metastasis in Vivo in Preclinical Models. A HeLa cells stably expressing either shCtrl or sh-circZFR (1 × 10⁶ cells) were injected into the footpads of mice and harvested popliteal lymph nodes (upper panel). Moreover, HeLa cells stably expressing either empty control or circZFR-OE (1 × 10⁶ cells) were injected into footpads of mice and harvested popliteal lymph nodes (lower panel). B Tumor volume analysis: Left panel shows comparison between empty vector and circZFR-OE groups; right panel shows comparison between shCtrl and sh-circZFR groups. C Representative images of harvested popliteal lymph nodes from the shCtrl, sh-circZFR, empty vector, and circZFR-OE groups. D Western blot analysis of FASN protein levels in popliteal lymph nodes from all four groups. Left: Representative Western blot images. Right: Quantification of FASN protein expression. E Schematic illustration of circZFR molecular mechanism. Overexpression circZFR binds to the m 6 A reader protein YTHDF3, facilitating m 6 A recognition on FASN mRNA and recruiting the translation initiator eIF4A3, boosting FASN translation, thereby facilitating EMT of cervical cancer cells. Statistical analyses of data in B and D were conducted using Student’s t-test. Data are presented as mean ± SEM from independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Cell lysates were incubated with beads conjugated to YTHDF3 or eIF4A3 antibodies (Proteintech).

Techniques: In Vivo, Stable Transfection, Expressing, Injection, Control, Comparison, Plasmid Preparation, Western Blot, Over Expression