Fig 1: Size distribution of nuclear puncta AGli31–455‐positive bodies are not Cajal bodies, nucleoli, polycomb bodies, or PML bodies. NIH 3T3 cells were transfected with GFP‐Gli31–455 and stained with antibodies against coilin (a Cajal marker), B23 (a nucleolus marker), CBX8 (a polycomb marker), or PML (a PML body marker).B NIH 3T3 cells were transfected with a construct expressing SC‐35‐GFP and GFP fluorescence was monitored in live cells. Snapshots at indicated time points show a nuclear speckle fusion event.C NIH 3T3 cells were transfected with a construct expressing GFP‐Gli31–455, and GFP fluorescence was monitored in live cells. Individual nuclear bodies were photobleached, and FRAP was monitored for 1 min. Data were normalized to the maximum and minimum intensity. The mean characteristic recovery time is indicated ± SEM.DHistograms depicting the size distribution of nuclear body areas are shown for cells transfected with HA‐SPOP alone and GFP‐Gli31–455 + HA‐SPOP.E–G NIH 3T3 cells were transfected with only HA‐SPOP or GFP‐Gli31–455 + HA‐SPOP. Box plots of the median aspect ratio of the (E) GFP‐Gli31–455‐positive nuclear bodies in one cell, (F) of the intracellular median area of nuclear speckles, and (G) of the number of bodies per cell for cells transfected with HA‐SPOP alone and GFP‐Gli31–455 + HA‐SPOP are shown. All three were significantly different (P = 1.2 × 10−42, P = 4.6 × 10−19, and P = 8.7 × 10−6, respectively, according to the Wilcoxon rank‐sum test), consistent with the different nature of distinct nuclear bodies. The medians are indicated as black horizontal lines within the boxes, and boxes enclose values between the first and third quartile. Interquartile range (IQR) is calculated by subtracting the first quartile from the third quartile. All values that lay more than 1.5× IQR lower than the first quartile or 1.5× higher than the third quartile are outliers that are plotted as squares. The smallest and highest values that are not outliers are connected with the dashed line. Source data are available online for this figure.