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. 2019 Mar 29;27(4):475–486. doi: 10.3727/096504018X15270916676926

Figure 2.

Figure 2

Bufalin affects cell activity in glioma spheroid cultures. (A) U87MG and LN-229 cells were cultured in two-dimensional (2D) and 3D (spheroid) cultures. The cells formed spheroids following a 10-day culture in serum-free medium. Scale bars: 100 μm. (B) The human embryonic stem cell markers Nanog, SOX2, and Oct4 were upregulated in 3D cell cultures compared with those in 2D cell cultures. (C) The cells formed spheroids following a 7-day culture in serum-free medium with bufalin (0, 5, and 20 nM in U87MG and 0, 10, and 40 nM in LN-229 cells). Scale bars: 100 μm. **p < 0.01, ***p < 0.001. (D) Cell spheroids were treated with bufalin (0, 10, and 20 nM in U87MG and 0, 40, and 80 nM in LN-229 cells) for 5 days. The number of spheroids (diameter ≥100 μm) was counted, and the results are expressed as the mean ± SD. Scale bars: 100 μm. ***p < 0.001. (E) U87MG and LN-229 cell spheroids were treated with bufalin (0, 5, 10, and 40 nM in U87MG and 0, 20, 80, and 160 nM in LN-229 cells) for 3 days and then stained with propidium iodide. All experiments were repeated three times. Scale bars: 50 μm.