Fig. 3: circARHGAP10 destabilizing MAT2A through the direct interaction with MAT2A.

A RIP assays were performed with an anti-AGO2 antibody in T24-CR cells to analyze circARHGAP10 enrichment. ***p < 0.001 versus input group. (B) Analysis for lineal and circular ARHGAP10 enrichment. RNA pull down was performed with sense probe which targeting the back splice junction site of circARHGAP10. ***p < 0.001 versus anti-sense group. C Sense probe and antisense probe were incubated with cell lysates of T24-CR -LV-circARHGAP10 cells for silver staining. D The peptide segment diagram of MAT2A was identified by mass spectrometry from RNA pulldown. E Sequence of identical peptides of MAT2A identified by mass spectrometry. F Co-localization of circARHGAP10 with MAT2A in BCa cells. Scale bar = 10 μm. G The interaction between MAT2A and circARHGAP10. H Protein levels of MAT2A in BCa cells with circARHGAP10 overexpression. I Protein levels of MAT2A in circARHGAP10 overexpressed BCa cells with MG132 (20 μM) treatment for 10 h. J Relative protein levels of MAT2A and GAPDH in BCa cells treated with 200 μM cycloheximide (CHX) at different time points. K Co-IP experiment was performed to determine the ubiquitination modification level of MAT2A in BCa cells. L Co-IP experiment was performed to determine the ubiquitination modification level of MAT2A in 293 T cells. M Relative expression of MAT2A and circARHGAP10. N The contingency of the correlation of the staining intensity of MAT2A and circARHGAP10 analyzed with chi-squared test.