Fig. 6

Alteration of PolD1 expression affects Golgi organization. a RPE1 cells were transfected with control or PolD1 siRNAs. Anti-α-tubulin fluorescence intensities were measured at the Golgi area, and the background intensity was obtained from cytoplasmic areas lacking microtubules. Here, the average microtubule intensities per pixel at the Golgi are presented after subtraction of the background. The Golgi area was outlined according to TGN46 staining by using the freehand selection option in ZEN software. Data are shown as means ± s.d. of three experiments; **p < 0.01, *p < 0.05, two-tailed, unpaired student’s t-test; control, n = 60 cells; PolD1 depletion, n = 50 cells per experiment. b Cells were transfected with GFP or GFP-PolD1 and then immunostained for GM130 and GCP6; n = 50 cells from three independent experiments. c RPE1 cells transfected with control or PolD1 siRNAs were treated with nocodazole. After nocodazole washout, Golgi reassembly was examined at the indicated time points. TGN46 immunofluorescence was used to measure the size of the Golgi particles that were not clustered in the Golgi region surrounding the centrosome. Quantification graphs show the average size of the individual particles and the total area of the particles. Data are presented as means ± s.e.m. of three experiments (30 cells analyzed per condition). Scale bars, 10 μm