Fig. 4: circARHGAP10 act as a scaffold for the interaction between TRIM25 and MAT2A to facilitate ubiquitination of MAT2A.

A Intersection of the proteomics between T24 and T24-CR cells and the mass spectrometry analysis of circARHGAP10 RNA-pulldown. B Differential protein levels of ISG15, USP5, USP14 and TRIM25 of T24 and T24-CR cells identified with proteomics analysis. C Binding of circARHGAP10 with TRIM25 in BCa cells. D Co-localization of TRIM25 with MAT2A in BCa cells. Scale bar = 10 μm. E Co-IP validated TRIM25 binding with MAT2A in T24-CR cells. F Molecular docking of the protein structure of TRIM25 and MAT2A. G MAT2A protein expression in BCa cells with knockdown of TRIM25. H Protein levels of MAT2A in BCa cells with the over-expression of TRIM25 or TRIM25ΔRBD. I Co-IP experiment was performed to identify the ubiquitination modification level of MAT2A in BCa cells with the over-expression of TRIM25 or TRIM25ΔRBD. J Co-IP experiment was performed to identify the ubiquitination level of MAT2A in 293 T cells with the over-expression of TRIM25 or TRIM25ΔRBD. K Co-IP experiment was performed to identify the ubiquitination level of MAT2A in 293 T cells with the over-expression of HA-Ubiquitin or the indicated mutants. L Co-IP experiment was performed to identify the ubiquitination level of MAT2A in BCa cells with the over-expression of TRIM25 or the silencing of circARHGAP10. M The protein level of MAT2A in BCa cells with the over-expression of TRIM25 or the silencing of circARHGAP10. N Co-IP experiment was performed to validate the correlation of circARHGAP10 with binding of TRIM25 with MAT2A in T24-CR cells. RNase A (10 μg/ml) and RNase R (100 U/ml) were the indicated treatment concentration.