FIGURE 1.
The cytotoxicity and apoptosis effect of imipramine on OS cells. (A) U‐2 OS and MG 63 cells treated with 0–100 μM imipramine for 24 and 48 h and assayed by MTT. (B) Colony formation of U‐2 OS and MG 63 cells after pre‐treated with different concentrations of imipramine for 48 h. (C) The staining of annexin‐V and PI after imipramine treatment for 48 h was assayed by flow cytometry on OS cells. (D) The protein expression of cleaved caspase‐3 and MCL‐1 after imipramine treatment for 48 h was assayed by Western blotting. (E) The activation of cleaved caspase‐3 after imipramine treatment for 48 h was determined by flow cytometry. (a vs. 0 μM 24 h; b vs. 0 μM 48 h; 2 p < 0.001; 3 p < 0.0005; 4 p < 0.0001). MTT assays and flow cytometry analyses were performed in five independent replicates, while Western blotting experiments were conducted in triplicate.