Fig. 7

The expression levels of p16 (A), p21 (B), p27 (C), and p53 (D) proteins in HepG2-derived spheroids following 24 h of treatment with Protodioscin (PD) and Methotrexate (MTX) were analyzed using the Western blot method. Control (control group), MTX (6 μm), MTX + PD 1.6 (6 μm MTX + 1.6µM PD), MTX + PD 4 (6 μm MTX + 4µM PD) and MTX + PD 8.14 (6 μm MTX + 8.14µM PD).GAPDH protein was used as a calibrator. The results are reported as mean ± standard deviation for tree replicates. *, **, *** and **** demonstrate significant difference between groups at P < 0.05, P < 0.01, p < 0.001 and p < 0.0001.