Figure 5. Glucose deprivation does not alter HNE-induced cell death. Differentiated SH-SY5Y cells were cultured in medium with 25 mM, 5 mM or 0 mM glucose, or 20 mM 2DG, for 24 h, followed by 30 μM HNE in the presence or absence of 40 μM CQ for 2 h. (A) Comparison of cell viability in response to lower glucose concentration vs. 20 mM 2DG for 24 h, followed by 30 μM HNE for 2 h. (B) Western blot analyses of protein extracts from cells treated with glucose-free medium or 2DG for SQSTM1, MAP1LC3A-I, and MAP1LC3A-II. ACTB was used as a protein loading control. (C) Quantification of MAP1LC3A-II from (B). Data represent mean ± SEM (n = 3), normalized to 0 mM glucose+CQ. *P < 0.05, the Student t test. (D) Quantification of SQSTM1 from (B). Data represent mean ± SEM (n = 3), normalized to 2DG. *P < 0.05, the Student t test.