Fig. 4.

In 6CFSMEo- cells, UPF1 protein is concentrated partially or totally in P-bodies after cytoskeleton inhibitor treatment. Cells were transfected with constructs expressing YFP–UPF1 and GPx1 Ter and then incubated with DMSO, cytochalasin D (CytoD), JPK, colchicine (COL) or Taxotere (TAX) for 48 h. After fixation and permeabilization, the cells were incubated first with anti-DCP1a primary antibody and then with the Alexa Fluor 594-conjugated secondary antibody (red). Finally, the nuclei were visualized in blue by Hoechst 33342 staining. Green arrowheads indicate UPF1 cytoplasmic foci; red arrowheads indicate the P-body marker DCP1a; orange arrowheads indicate colocalization of UPF1 and DCP1a. The histogram represents the percentage of colocalization between UPF1 foci and DCP1a foci. Cells (mean±s.d.; n=10) from three different experiments were counted for each condition.