Journal: bioRxiv

Article Title: Resistance to Radiation Enhances Metastasis by Altering RNA Metabolism

doi: 10.1101/2025.02.19.638943

Figure Lengend Snippet: A. The Pearson correlation of the circular RNA isoforms for each biological replicate of the HNRNPL knockdown cells and RR cells. B. The expression of the ceRNAs based on RT-qPCR in T47D cells given empty vector or HNRNPL-FLAG plasmid. C. The expression of circRAB12 , circBACE2 , and circBIRC6 in the control cells (siNC) compared to the siRNA knockdown cells (si-ceRNA-1 and si-ceRNA-2). D. The mRNA expression of circRAB12 after transfection of the circRAB12 or empty plasmid into the HNRNPL knockdown cells. The mRNA expression of ITGβ3 in those cells. Data are represented as mean ± SD. The P values in panel B and D were obtained unpaired t-test two-tailed, in panel C were obtained by one-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: The circular RNA forming plasmid was digested with BsmbI-v2 (R0739S, NEB) and then cloned with the circRAB12 gBlock using the 2x HiFi DNA Assembly Master mix (E2621S) based on the manufacturer’s instructions.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Plasmid Preparation, Control, Transfection, Two Tailed Test

Journal: bioRxiv

Article Title: Resistance to Radiation Enhances Metastasis by Altering RNA Metabolism

doi: 10.1101/2025.02.19.638943

Figure Lengend Snippet: A. IMCR-seq results showing the differential expression of different circular RNAs in the MDA-MB-468-RR and in the shHNRNPL-1 cells. The blue dots highlight the top 5 differentially expressed circular RNAs whereas the other labels represent other circular RNAs that can function as ceRNAs B. A graph showing the let-7 binding score for each circular RNA in comparison to its abundance (based on ranking of circular RNAs from CPM values) in MDA-MB-468-RR cells. C. mRNA expression of the top ceRNAs identified by Circr in the MDA-MB-468-RR and HNRNPL-knockdown cells (n=3, mean ± SD, one-way ANOVA with Dunnett’s multiple comparisons tests). D. mRNA expression of the top ceRNAs in MDA-MB-468 transfected with empty vector and HNRNPL (n=3, mean ± SD, unpaired t-test two-tailed). E. mRNA expression of ITGβ3 after siRNA treatment of the ceRNAs, with a representative image of the surface expression of integrin β3 and quantification based on flow cytometry (n=3, mean ± SD, unpaired t-test two-tailed). F. Representative flow cytometry data showing the surface expression of integrin β3 in the HNRNPL knockdown cells transducing with an empty plasmid or circRAB12 . The graphs quantify the fold change in integrin β3 surface expression in the HNRNPL knockdown cells after transducing them with an empty plasmid or circRAB12. n=3, mean ± SD, unpaired t-test two-tailed.

Article Snippet: The circular RNA forming plasmid was digested with BsmbI-v2 (R0739S, NEB) and then cloned with the circRAB12 gBlock using the 2x HiFi DNA Assembly Master mix (E2621S) based on the manufacturer’s instructions.

Techniques: Expressing, Binding Assay, Comparison, Knockdown, Transfection, Plasmid Preparation, Two Tailed Test, Flow Cytometry