Figure 3
From: Effect of HIV-1 Tat on the formation of the mitotic spindle by interaction with ribosomal protein S3
Knockdown of RPS3 results in aberrant mitotic spindle formation, segregation failure, and defective abscission. (A) Aberrant mitotic spindle formation and segregation failure in RPS3-depleted cells. HeLa cells were transduced with shRPS3-4 and stained with anti-α-tubulin and anti-RPS3 antibodies. Control, n = 71; shRPS3, n = 112. Experiments were repeated three times. (B) Transduction of shRPS3 resulted in a defect in abscission. HeLa cells were transduced with shRPS3-4 and stained with an anti-α-tubulin antibody. (C) The average length of bridges between two daughter cells is plotted for control and RPS3-knockdown cells. Data represent the mean ± SEM of three independent experiments. Statistical analysis was performed using the two-tailed t test. Control, n = 356; RPS3-knockdown, n = 356. P = 0.0008. (D) Time-lapse images of control and shRPS3-4-transduced HeLa-H2B-GFP cells. HeLa-H2B-GFP cells were transduced with the control or shRPS3-4 virus and incubated for 2 days. Cells were plated onto chamber slides and incubated for a further 24 h. Images were acquired using a 40×, NA 0.6 dry objective lens every 5 min in a chamber at 37 °C in 5% CO2 and processed using ImageJ software.
