Fig. 2. ALDH⁺ CD133⁺ CCSCs are IR resistant and IR induces CCSCs in a dose dependent manner.

Cells were seeded (1 × 10⁵ cells/2 mL culture medium/well) in 6-well plates and incubated for 24 h, followed by IR with 8Gy, 10Gy, 12Gy, and either cultured for an additional 24 h or IR with 10Gy and cultured for an additional 24 h, 48 h, or 72 h. The treated cells were harvested and analyzed for ALDH⁺CD133⁺ cells. The total number of cells was counted using the Trypan blue exclusion method. ALDH⁺CD133⁺ cells were calculated based on the total cell number and percentage of ALDH⁺CD133⁺ cells detected by flow cytometry (A, C). The mean ± SD of absolute cell numbers of total cells and ALDH⁺CD133⁺ cells are shown (B, D). * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.