Figure 2. Depletion of 4E-BP1 leads to increased outcome of polyploidy, aberrant spindles and misaligned chromosomal DNA in normal cultured cells. (A) Forty-eight h after transfection with anti-4E-BP1-specific and non-specific siRNA, HepG2 cells were fixed and co-stained with antibodies against α-tubulin and γ-tubulin. Mitotic cells with abnormal spindles were captured. Representative images indicate the misaligned chromosomal DNA (unattached free DNA) and multipolar spindle or multiple centrosomes in 4E-BP1-depleted cells. (B) The polyploidy of the misaligned chromosomal cells to metaphase cells and multipolar spindle cells to mitosis cells were counted (* p < 0.05, ** p < 0.01 as compared with control cells). (C) Forty-eight h after transfected with anti-4E-BP1 specific and non-specific siRNA, HepG2 cells were collected and analyzed by flow cytometry. (D) Quantitative data of the polyploidy cells were presented as the mean and SD from three independent experiments (** p < 0.01 as compared with control cells).