Fig. 6

Effects of Equol and TRIM21 on the ubiquitination of AXNA2. A The PPI network showing the proteins interacted with TRIM21. B Double immunofluorescence staining of TRIM21 (red) and AXNA2 (green) in JEG-3 and Bewo cells. C Co-immunoprecipitation of TRIM21 with AXNA2 in JEG-3 and Bewo cells. D JEG-3 and Bewo cells were transfected with siTRIM21 or siNC for 48 h, and then treated with 40 μM Equol for 24 h. The expression of AXNA2 was measured by western blot. E JEG-3 and Bewo cells were transfected with siTRIM21 or siNC for 48 h, and 12-h treatment with MG132 (10 μM) was subjected before cell harvesting. F JEG-3 and Bewo cells were treated with 40 μM Equol for 24 h, and 12-h treatment with MG132 (10 μM) was subjected before cell harvesting. E and F The ubiquitination of AXNA2 was detected by co-immunoprecipitation of AXNA2 with ubiquitin. G JEG-3 and Bewo c ells were treated with 40 μM Equol and 10 μM MG132 for 24 h. Western blot analysis of AXNA2 was performed. Veh. Vehicle, Ub ubiquitin