Journal: Cell Death & Disease
Article Title: Circular RNA TFRC/ SCD1 mRNA interaction regulates ferroptosis and metastasis in gastric cancer
doi: 10.1038/s41419-025-07759-x
Figure Lengend Snippet: A Venn diagram showing the intersection of potential circTFRC-binding proteins predicted by ENCORI and RBPDB databases. B Representative Western blotting showing ELAVL1 enrichment upon circTFRC pulldown in GC cell lysates, with Actin as a negative control. C qRT–PCR analysis demonstrating circTFRC enrichment in an anti-ELAVL1 RIP assay in GC cells. IgG serves as a control. D Venn diagram displaying the intersection between ferroptosis-suppressor genes in the FerrDb database and ELAVL1-binding mRNAs predicted by ENCORI. E qRT–PCR analysis showing the expression of mRNAs in GC cells under control conditions (sh-NC) or circTFRC knockdown (sh-circTFRC). F qRT–PCR analysis showing the expression of mRNAs in GC cells transfected with control or ELAVL1 overexpression vector. G Western blotting showing SCD1 protein levels in GC cells under control conditions (sh-NC) or circTFRC knockdown (sh-circTFRC), with Actin as a loading control. H Western blotting of ELAVL1 and SCD1 in GC cells transfected with control vector, or ELAVL1 overexpression vector. I qRT–PCR analysis showing SCD1 3’ UTR enrichment in an anti-ELAVL1 RIP assay in GC cells, with CKAP5 mRNA as a positive control and GAPDH mRNA as a negative control. J qRT–PCR analysis of SCD1 mRNA stability in control, ELAVL1 -overexpressing, circTFRC knockdown, and ELAVL1- overexpressing + circTFRC knockdown cells following Actinomycin D treatment (5 µg/mL) at various time points (0, 1, 2, 4, and 8 h). K qRT–PCR demonstrating circTFRC and SCD1 3’ UTR enrichment upon circTFRC pulldown in GC cell lysates. GAPDH serves as a negative control. L qRT–PCR showing SCD1 3’ UTR enrichment after anti-ELAVL1 RIP in GC cells under control conditions (sh-NC) or circTFRC knockdown (sh-circTFRC). M Representative Western blot of ELAVL1 and SCD1 in GC cells after the transfection of the control vector or ELAVL1 overexpression vector or cotransfection of ELAVL1 + sh-circTFRC vectors. The data are shown as the mean ± SD. The P values were determined by a two-tailed unpaired Student’s t test ( C , E , F , I , K , L ) or two-tailed two-way ANOVA ( J ); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. See also Figs. and .
Article Snippet: Two siRNAs targeting circTFRC BSJ regions and a negative control siRNA (si-NC) were synthesized by GenePharma.
Techniques: Binding Assay, Western Blot, Negative Control, Quantitative RT-PCR, Control, Expressing, Knockdown, Transfection, Over Expression, Plasmid Preparation, Positive Control, Cotransfection, Two Tailed Test