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. 2018 Sep 1;11(9):4305–4318.

Figure 6.

Figure 6

Down-regulated BAG3 decreased HIF-1α protein expression through enhancing more formation of HSP70-HIF-1α complex and promoting degradation of HIF-1α by HSP70. A. Proteasome inhibitor increased the expression of BAG3, HIF-1α, and HSP70 in U87 glioblastoma cell line. With the treatment duration of proteasome inhibitor MG132 (2.5 μM) extended, the expression of BAG3, HIF-1α and HSP70 increased. B. Relative expression level of BAG3, HIF-1α, and HSP70 in U87 glioblastoma cell line. C. MG132 increased the expression of BAG3, HIF-1α, and HSP70 in U251 glioblastoma cell line. With the treatment duration of MG132 (2.5 μM) extended, the expression of BAG3, HIF-1α and HSP70 increased. D. Relative expression level of BAG3, HIF-1α, and HSP70 in U251 glioblastoma cell line. E. HSP70 inhibitor increased the expression of HIF-1α. With the concentration HSP70 inhibitor KNK437 increased, HIF-1α expression increased. F. Relative expression level of BAG3, HIF-1α, and HSP70 after a treatment of KNK437. G. Immunoprecipitation of U251 glioblastoma cell line. Whole protein was immunoprecipitated using BAG3, HIF-1α, HSP70 and IgG antibody, and immunoprecipitates were analyzed by western blot to detect BAG3, HIF-1α, and HSP70. Protein expression level was tested by western blot. Each experiment was repeated three times. Protein and mRNA expression level were normalized relative to the amount of GAPDH. Data were showed as means ± SD. *P < 0.05 represented statistical significance.