biotin-labeled tirna-met sense and antisense sequences  (GenScript corporation)

Journal: Cellular & Molecular Biology Letters

Article Title: The novel tRNA-derived fragment, tiRNA-Met, inhibits the malignant progression of triple-negative breast cancer by regulating RANBP3L via a targeted interaction with SNRPA

doi: 10.1186/s11658-025-00738-2

Figure Lengend Snippet: tiRNA-Met directly interacted with SNRPA and promoted its ubiquitination. A , B Silver-stained SDS-PAGE gel images showed proteins immunoprecipitated by tiRNA-Met and its antisense RNA in MDA-MB-231 ( A ) and BT-549 ( B ) cell lines. C Western blot analysis of products from RNA pull-down assays using tiRNA-Met sense and antisense confirmed that tiRNA-Met directly associated with SNRPA. D , E RIP followed by RT-qPCR demonstrated specific binding of tiRNA-Met to SNRPA in MDA-MB-231 ( D ) and BT-549 ( E ) cells. F RNA pull-down and western blot assays revealed that tiRNA-Met bound to the RRM2 domain of SNRPA. G The mRNA expression levels of SNRPA remained unchanged in MDA-MB-231 and BT-549 cells transfected with tiRNA-Met mimics or inhibitor. H Overexpression of tiRNA-Met reduced SNRPA protein levels in both cell lines, while inhibition of tiRNA-Met increased SNRPA protein levels. I MDA-MB-231 cells treated with cycloheximide (Chx; 5 μM) and then transfected with tiRNA-Met mimics exhibited a reduced half-life of endogenous SNRPA protein, as shown by western blot; β-actin served as a loading control. J Treatment with MG132 (20 μg/ml) led to a progressive increase in SNRPA protein levels over time; β-actin was used as a reference. K Immunoprecipitation assays indicated that the binding of SNRPA to ubiquitinated proteins was elevated in MDA-MB-231 cells transfected with tiRNA-Met mimics. All data are presented as means ± SD; ns, P > 0.05; * P < 0.05; ** P < 0.01; **** P < 0.0001

Article Snippet: Biotin-labeled tiRNA-Met sense and antisense sequences were obtained from Genescript (Nanjing, China).

Techniques: Ubiquitin Proteomics, Staining, SDS Page, Immunoprecipitation, Western Blot, Quantitative RT-PCR, Binding Assay, Expressing, Transfection, Over Expression, Inhibition, Control