Figure 6.

GOLPH3 expression is necessary for timely G1-phase progression. (A) RT-qPCR and Western analysis of U2OS cells transfected with non-target control siRNAs (siCTRL) or with siRNAs specific for GOLPH3 (siGOLPH3). mRNA expression values were normalized to the expression of EIF2C2, used as standard control. Data are represented as fold-change (mean + SD) relative to siRNA control. *, p < 0.05. GOLPH3 protein levels were assessed by Western analysis with a specific antibody. (B) U2OS cells were transfected with control or GOLPH3-specific siRNAs and subsequently synchronized in mitosis by Thy-Noc treatment. After release of cells into the cell cycle, samples were collected at several time-points, incubated with PI and cell cycle phases were assessed by FACS analysis. Data represent the mean ± SD of the fraction of siCTRL or siGOLPH3-transfected cells in G1 or S at each time-point obtained from three independent experiments. *, p < 0.05.