Fig. 6: SH3RF2 impaired the stability of RBPMS via its E3 ubiquitin ligase activity.

a 3D structure of SH3RF2 from the UniProt database (https://www.uniprot.org/). SH3RF2 contains a RING domain (12-52 aa) and exerts E3 ubiquitin ligase activities. b The PPI network analysis from the IntAct Molecular Interaction Database (https://www.ebi.ac.uk/intact/home) exhibits that SH3RF2 interacts with 25 proteins including RBPMS. c Pearson correlation analysis between IHC scores of SH3RF2 and RBPMS in the tumor tissues from OC patients. n = 120. d Colocalization of endogenous SH3RF2 and RBPMS was detected by immunofluorescence (Scale bars, 25 μm). n = 3. e Co-immunoprecipitation of SH3RF2 with RBPMS using anti-SH3RF2 and anti-RBPMS. Immunoblotting was performed with anti-RBPMS and anti-SH3RF2. n = 3. f SH3RF2 and RBPMS expressions in DDP-resistant OC cells with stable depletion of SH3RF2 were evaluated by western blotting. n = 3. g RBPMS expressions in the xenograft tumor tissues generated by DDP-resistant OC cells with stable depletion of SH3RF2 were evaluated by western blotting. n = 6. h DDP-resistant OC cells with stable depletion of SH3RF2 were treated with 20 µg/mL CHX or/and 5 μM MG132 for 0, 3, 6, and 9 h. Western blots of lysates from cells stained for RBPMS. n = 3. i Co-immunoprecipitation of RBPMS with K48 ubiquitin in DDP-resistant OC cells with stable depletion of SH3RF2 using anti-RBPMS. Immunoblotting was performed with anti-RBPMS and anti-K48 ubiquitin. n = 3. j Co-immunoprecipitation of RBPMS with K48 ubiquitin in DDP-resistant OC cells transfected with wild-type SH3RF2 or the RING mutant of SH3RF2 using anti-RBPMS. Immunoblotting was performed with anti-RBPMS and anti-K48 ubiquitin. n = 3. Data are expressed as the mean ± SD. The p values were determined by one-way or two-way ANOVA.