Fig. 3: UBE2V1 favors ELVAs formation.
From: UBE2V1 governs aging induced protein aggregation and developmental defects in oocytes and embryos
A Schematic diagram of the UBE2V1 overexpression and knockdown experiment in oocytes. B Validation of exogenous Myc-UBE2V1 overexpression and UBE2V1 knockdown efficiency in oocytes. Western blot analysis was performed using anti-UBE2V1 antibody to confirm exogenous Myc-UBE2V1 mRNA overexpression and siRNA-mediated UBE2V1 depletion in GV oocytes. Tubulin was used as a loading control. Protein lysates from 100 GV oocytes were loaded per lane. C Confocal images of control (7–8 weeks), UBE2V1 overexpression (UBE2V1-OE), and UBE2V1 knockdown (UBE2V1-KD) oocytes at the GV stages labeled with Proteostat (red). D Quantification of the number of the Proteostat compartments. E Western blot analysis of K63 poly-ubiquitination protein levels in Young (7–8 weeks), Old (42–45 weeks), and Young with UBE2V1 overexpression oocytes. Tubulin was used as a loading control. Total proteins from 80 oocytes were loaded in each lane. F Western blot analysis was performed using anti-Myc antibody to confirm the overexpression of Myc-UBE2D3 mRNA. G Confocal images of control (7–8 weeks) and UBE2D3-overexpressing (UBE2D3-OE) oocytes at the GV stage labeled with Proteostat (red) and counterstained with Hoechst 33342 for nuclear DNA visualization (blue). H Quantification of the number of the Proteostat compartments. I Western blotting analysis of K63 polyubiquitinated protein in UBE2D3-OE, control and UBE2V1-OE oocytes. Tubulin was used as a loading control. Protein lysates from 90 GV oocytes were loaded per lane. The total number of oocytes analyzed is indicated in the figures. Data were presented as mean ± SD. Statistical analysis was performed using an unpaired two-tailed Student’s t-test for (H), and one-way ANOVA followed by multiple comparisons for (D). A P-value < 0.05 was considered statistically significant.
