Fig. 7: 4-MU inhibits glioma growth in vivo and, when combined with autophagy inhibitors, exerts synergistic effects on glioma cell viability, autophagy levels, and the cell cycle.

A Viability of U251 glioma cells cultured with 4-MU, followed by treatment with CQ (30 μmol/L) for 48 h. B Levels of the Ki67 protein in U251 glioma cells were detected using immunofluorescence staining after culture with 4-MU, followed by treatment with CQ (30 μmol/L) for 48 h. C Relative levels of the P62 and MAP1LC3B proteins in U251 glioma cells cultured with 4-MU, followed by treatment with CQ (30 μmol/L) for 48 h. Scale bar: 50 μm. D The cell cycle distribution was detected in U251 glioma cells using flow cytometry after culture with 4-MU, followed by treatment with CQ (30 μmol/L) for 48 h (green: G0-G1, yellow: S, and blue: G2-M). E Relative levels of the CCNB1 and CCND1 proteins in U251 glioma cells cultured with 4-MU, followed by treatment with CQ (30 μmol/L) for 48 h. F Representative images of IHC staining for Ki67 and HA in the orthotopic xenograft tumors from the control and 4-MU treatment groups. Scale bar: 50 μm. G–H Representative MRIs of orthotopic xenograft tumors and survival curves of the control and 4-MU treatment groups. P: P-value for the comparison of the control and 4-MU groups. The data are presented as the mean ± SD; *P < 0.05, **P < 0.01, and ***P < 0.001, ****P < 0.0001.