Fig. 4: Overexpression of Akt2, as a major negative regulator of the FoxO3a activity, mediates GC resistance in lymphocytes.

a Western blot analysis of Akt1 and Akt2 expression in CCRF-CEM cells, CEM-DR cells, Jurakt cells, Daudi cells, and L-02 cells. b Western blot analysis of Akt1 and Akt2 expression in CCRF-CEM cells with increasing of IC50 value of DEX. GAPDH was used as a loading control. c Real-time quantitative PCR analysis of Akt1 and Akt2 mRNA in lymphoctyes from bone marrow of newly diagnosed or relapse/refractory ALL patients. d Diagnostic sensitivity and specificity analysis of Akt2 mRNA as an indicator of GC resistance in ALL patients. e Western blot analysis of Akt1/Akt2 and MYC after FoxO3a immunoprecipitation in 293T cells expressing Flag-tagged Akt1/Akt2 and MYC-tagged FoxO3a. f FoxO3a western blot analysis after Akt1/Akt2 immunoprecipitation in 293T cells expressing Flag-tagged Akt1/Akt2 and MYC-tagged NR3C1. g Western blot analysis of Akt1 or Akt2 after FoxO3a protein immunoprecipitation in Jurkat cells. h Western blot analysis of Akt1 and Akt2 expression in Jurkat T-ALL cells after Akt1 iRNA or Akt2 iRNA transfection. i Analysis of apoptosis in Jurkat T-ALL cells after Akt1 iRNA or Akt2 iRNA transfection with 24 h incubation with DEX. j Western blot analysis of FoxO3a and p-FoxO3a (Ser253) expressions in Jurkat T-ALL cells after Akt1 iRNA or Akt2 iRNA transfection with 24 h incubation with DEX. k Western blot analysis of Akt1 and Akt2 expression in Jurkat T-ALL cells after Akt1 plasmid or Akt2 plasmid transfection. l Analysis of apoptosis in Jurkat T-ALL cells after Akt1 plasmid or Akt2 plasmid transfection with 24 h incubation with DEX. m Western blot analysis of FoxO3a and p-FoxO3a (Ser253) expressions in Jurkat T-ALL cells after Akt1 plasmid or Akt2 plasmid transfection with 24 h incubation with DEX. Bar graphs in c, i, l represent mean ± SD