Figure 3.

Endogenous TRIM5α Co-localizes with Cajal Bodies in Human and RM DCs

(A and B) Co-localization of TRIM5α with (A) PML-NB and (B) the Cajal body marker Coilin in human PBLs and DCs ± 100 U IFNα for 24 hr, 5 or 50 μg/ml leptomycin B (LMB) for 4 hr. Scale bars represent 5 μm. Co-localization was assessed using the Image J plugin Just Another Colocalization Plugin (JACoP) with threshold set at 20 for all images in both fluorescence channels. Graph shows Manders’ coefficients for the fraction of PML-NB overlapping with TRIM5α, mean of three independent experiments ± SEM. In total, a minimum of ∼50 cells were analyzed for each condition.

(C) Proximity between TRIM5α and coilin was assessed by Duolink proximity ligation assay (red dots). Graph shows the average number of spots over the number of Hoechst-stained nuclei in eight random fields per cell type ± SEM (Photoshop CS5).

Scale bars represent 5 μm. Statistical significance was assessed by non-parametric unpaired t test (^∗∗∗p ≤ 0.001; ^∗p ≤ 0.05; ns, non-significant).