Fig. 2

Nutlin-3 stabilizes MDM2-CD276 protein-protein interactions in situ. a Proximity ligation assay of A375 cells treated with DMSO or 10 μM Nutlin-3 for 24 h. Green fluorescence spots suggest that both target proteins (MDM2-CD276) are within interacting range. The cells were counterstained with DAPI (blue). b Quantitation of CD276-MDM2 interaction after incubation with DMSO or Nutlin-3. The results represent the mean ± SD of technical triplicates. Statistical analysis was performed by one-way ANOVA with a Bonferroni post hoc test, *** p < 0.001. c Western blot of A375 cells treated with the indicated concentrations of Nutlin-3 (Nut-3). MDM2, PD-L1, CD276 and β-actin have apparent molecular weights of 90, 50, 60 and 42 kDa, respectively