Fig. 3.

PolD1 overexpression inhibits Golgi-associated microtubule regrowth. a, b RPE1 cells were transfected with GFP or GFP-PolD1 constructs and then subject to cold-induced microtubule depolymerization, after which microtubule regrowth was performed and examined through immunostaining. Hoechst 33258 was used to stain DNA. In the cells transfected with the PolD1 mutants D515V and S605del, microtubule regrowth was for 1.5 min (b). The images shown represent the phenotypes identified from three experiments (100 cells were analyzed per sample, and cells expressing GFP, GFP-PolD1, or PolD1 mutants at similar levels were selected for analysis). Scale bars, 10 μm. c Golgi-associated microtubules at 1.5 min of regrowth were quantified. The quantified data were normalized by the Golgi mass, and are presented as means ± s.d. from three independent experiments; ***p < 0.001, two-tailed, unpaired student’s t-test