Figure 8.

Neither nutlin-3a nor MEG3 significantly affected Mdm2–p53 binding.MEG3 was induced by treatment of U2OS-MEG3 cells for 20 h with or without 1 μg/ml doxycycline and/or 10 μM nutlin-3a. Cells were then fixed and stained for 2-color dSTORM of p53 and Mdm2. For each condition, single molecule localizations were collected from 10 randomly chosen cells in three separate experiments. A, fraction of pairs associated, as defined by a probability of chance association <0.1 (i.e., correction for local density) and distance <200 nm (upper limit for binding distance, accounting for error). B, median distance between pairs for each cell (nm). Boxes indicate median ± upper and lower quartile; whiskers indicate the range excluding outliers. Data points are colored by replicate. Means for each replicate are indicated by same-colored squares. ∗ indicates p < 0.05 by ANOVA. dSTORM, direct stochastic optical reconstruction microscopy.