Figure 5
From: The role of autophagy in asparaginase-induced immune suppression of macrophages
Asparaginase downregulates autophagy by activating Akt/mTOR and inhibiting Erk1/2 signaling pathway. (a–d) Ana-1 and RAW264.7 cells were treated with 300 IU/ml IFN-γ and 200 ng/ml LPS, either alone or in combination with 0.1 IU/ml asparaginase for 24 h. (a) The expression level of mTOR, p-mTOR, Akt, p-Akt, p70s6K, p-p70s6K, 4EBP1, and p-4EBP1 in Ana-1 cells were analyzed by western blot. Quantification of p-mTOR/mTOR, p-Akt/Akt, p-p70s6K/p70s6K, and p-4EBP1/4EBP1 ratios in different groups were presented in bar graphs. (b) The expression of mTOR, p-mTOR, Akt, p-Akt, p70s6K, p-p70s6K, 4EBP1, and p-4EBP1 in RAW264.7 cells were analyzed by western blot. Quantification of p-mTOR/mTOR, p-Akt/Akt, p-p70s6K/p70s6K, and p-4EBP1/4EBP1 ratios in different groups were presented in bar graphs. (c) Western blot was performed to analyze the protein Erk1/2 and p-Erk1/2 in Ana-1 cells. Quantification of p-Erk1/2/Erk1/2 ratios in different groups were presented in bar graphs. (d) Western blot was performed to analyze the protein Erk1/2 and p-Erk1/2 in RAW264.7 cells. Quantification of p-Erk1/2/Erk1/2 ratios in different groups were presented in bar graphs. Densitometric values were quantified using the ImageJ software and normalized to control. The values of control were set to 1. Results were represented as mean±S.D. (*P<0.05, **P<0.01)
