Figure 4.

ATP1A1 knockdown in GSCs induces cell-cycle arrest and increases apoptosis. (A) The cellular DNA content was analyzed by flow cytometry. The histogram shows the percentage of cells in each phase of the cell cycle (right panel). The data represent means of five experiments. (B) Immunoblot analysis of key proteins in the G1, S, and G2–M cell-cycle phases in sh-ATP1A1- and sh-NC-transfected GSCs. (C) FACS analysis of cell apoptosis in sh-ATP1A1- and sh-NC-transfected GSCs. The cells were double stained with Annexin V and PI. Early apoptotic cells are in the lower right quadrant, late apoptotic cells are in the upper right quadrant. The histogram in the right panel shows average percentages of apoptotic cells which contain the early apoptotic cells and late apoptotic cells. (D) Immunoblot analysis of apoptosis-related proteins in sh-ATP1A1- and sh-NC-transfected cells. Experiments were repeated independently at least three times (A–D). ^*P < 0.05, ^**P < 0.01.