Fig. 5

ZC3H8 and PML localization is altered by CK2 inhibitor TBB. a) cV1A 03–31 cells were treated with 10 μM of the CK2 inhibitor TBB for 2 h and processed for immunofluorescence using antibodies against PML and ZC3H8. Control cells were treated with DMSO only, and washout cells had the TBB media replaced with fresh media for 2 h. b) Quantification of the number of nuclear foci stained with PML and ZC3H8 (PML Bodies) treated with DMSO, TBB, or TBB and washout. Differences between control and TBB washout were not significant, but both were significantly different from TBB treated cells, p < 0.01. c) Cells transiently transfected with vector encoding ZC3H8-WT, ZC3H8-T32A, or ZC3H8-T32E phosphorylation mutants and processed for immunofluorescence against PML (red) and V5 tag (green). d) Quantification of PML-stained nuclear bodies in cells transfected with Zc3h8 phosphorylation mutants. Differences between vector, WT, and T32A cells were not significant, but all differed from the T32E mutant. Experiments were performed in triplicate, error bars represent standard deviation, and p values were determined by Tukey’s post hoc at p < 0.01., Images are representative of typical results. In low magnification images, scale bars represent 20 μm, and represent 5 μm in high magnification images