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. 2021 Apr 3;11(12):6006–6018. doi: 10.7150/thno.59841

Figure 5.

Figure 5

3-HAA-upregulated DUSP6 also diminishes Akt activity by dephosphorylating PDK1. A. DUSP6 expression detection in SMMC7721 cells treated with 3-HAA for 24 h at the indicated dose. B. The effect of DUSP6 on the Akt activity. T308 phosphorylation of Akt was detected in SMMC7721 cells overexpressing DUSP6 or depleted of DUSP6. C. Detection of DUSP6 binding proteins by mass spectrometry following co-immunoprecipitation. D. Detection of DUSP6 in PDK1-associated proteins. PDK1 was tagged with flag. E. The effect of DUSP6 overexpression on PDK1 and Akt activity. PDK1 phosphorylation (S241) and AKT phosphorylation (T308) were detected separately. F. DUSP6 knockdown fully restored 3-HAA-reduced PDK1 phosphorylation (S241) and partially recovered 3-HAA-deminished AKT phosphorylation (T308). The SMMC7721 cells were treated with 100 μM of 3-HAA for 24 h. G. PDK1 activator (PS210) rescued 3-HAA-induced cell death. The dose of PS210 was 10 μM. All cells were treated with 5 μM sorafenib for 4 days. The histogram shows the relative viability of HCC cells treated with 100 μM of 3HAA with or without 10 μM PS210.