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Fig. 4 | Cancer Cell International

Fig. 4

From: Extracellular HMGB1 interacts with RAGE and promotes chemoresistance in acute leukemia cells

Fig. 4

The ablation of RAGE enhanced apoptosis in leukemia cells and the apoptosis was related with the phosphorylated of P53. A (a-1) Cells that stably transfected with NC shRNA and RAGE shRNA were pretreated with 50 ng/mL rHMGB1 for 24 h, then cells were co-cultured with 0.4 μM ADM for another 24 h. Western blotting was used to detect the alternation of apoptosis. (a-2) Was the quantitative data of cleaved caspase 3/GAPDH. B (b-1) Cells that stably transfected with shRNA-RAGE were pretreated with the small inhibitor of p53, Pifithrin-β (1 μM) for 12 h, following the treatment of ADM (0.4 μM). Cells were subjected to western blotting to assess the level of apoptosis and the phosphorylation of P53. (b-2) Were the quantitative data of Bcl 2/Bax and PUMA/GAPDH in both leukemia cells. C Cells in step A were collected and washed with PBS for 2 times. Annexin V APC-A and APC-Cy7-A were used to evaluate the level of apoptosis in different groups. D Leukemia cells were treated with ADM (0.8 μM) with or without FPS-ZM1 for 24 h, Western blotting was used to detect the alternation of apoptosis. Data are the mean ± standard deviation of three independent experiments. *P < 0.05, **P < 0.01, compared with the ADM-treated NC group

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