Fig. 3

The role of the potential compounds in the self-renewal of P19 cells. A Effect of MDLS on the plate colony formation of P19 cells. P19 cells were plated onto 6-well plates for 24 h and then the culture medium was replaced with the fresh culture medium containing 5 μg/mL MDLS daily. After one week, the cells were stained with MTT and the colony formation was determined by the microscopic examination. B Effect of MDLS on the colony formation of P19 cells in hanging drop culture. P19 cells were made into a single-cell suspension and then cell suspension was added with 5 μg/mL of MDLS or DMSO. After that, the cells were counted and prepared at a density of 1 × 10⁴ cells/mL. Then, cells were placed on the underside of a petri dish lid in a 25 μl drop. The lid was inverted and placed on a plate containing 3 mL PBS. The morphological changes of the cells were observed under the microscope after 48 h. C Effect of MDLS on the colony formation of P19 cells in suspension culture. P19 cells were plated onto 0.5% agarose-coated six-well plates and treated with MDLS (5 μg/mL) or DMSO for 5 days. Subsequently, we harvested the embryoid bodies and transferred them to another 6-well plate. After adhering to the wall, the number of EBs was counted and the cells were stained with MTT