Fig. 3.

IFT172 and KIF3A siRNA knockdown promotes cell proliferation and cell cycling in confluent MSCs. Mesenchymal stem cells were cultured for 6 days and treated with siRNA on D1 D3 and D5. MSCs were collected on D0, D1, D2, D3, D4, D5 and D6, and the DNA content of proliferating MSCs was quantified by CyQUANT. Mitotic MSCs under confluent conditions (150,000 cells per well in 12 wells for 6 days) were stained for phosphorylated histone H3. For the cell cycle analysis, MSCs were cultured to confluence (at a density of 150,000 per well in a 12-well plate and cultured for 6 days, and treated with siRNA three times on D1 D3 and D5). MSCs were serum starved for 24 h and then passaged at a 1:3 ratio in standard medium containing serum. MSCs were collected and fixed at 0 h, 12 h and 24 h. MSCs were stained with propidium iodide, and the cell cycle was analyzed with a Beckman Coulter Cyan ADP flow cytometer