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. 2023 Mar 15;13(6):2701–2714. doi: 10.1016/j.apsb.2023.03.011

Figure 4.

Figure 4

HMGB2 was required for phosphatidylinositol 3-kinase III activity and autophagy induction. (A–C) Wildtype (WT) N2a cells and Hmgb2 knockout (KO) N2a cells were treated with 20 μmol/L corynoxine B (Cory B) for 6 h. Expression levels of LC3II and p62 were assessed by Western blot, and quantifications are shown. (D, E) RFP-GFP-LC3-transfected N2a cells and Hmgb2 KO N2a cells were untreated or treated with Earle's balanced salt solution (EBSS) for 6 h. Fluorescence was observed under a confocal microscope, and representative figures are shown. Quantification of LC3 puncta numbers is shown. n = 30 cells per group. (F–H) WT N2a cells and Hmgb2 KO N2a cells were treated with EBSS or 1 μmol/L Torin 1 alone or cotreated with chloroquine (CQ) for 6 h. Expression level of LC3II was assessed by Western blot, and quantifications are shown. (I–K) EGFP-2xFYVE and RFP-LC3 co-transfected N2a cells or Hmgb2 KO N2a cells were untreated or treated with EBSS or 1 μmol/L Torin1 for 6 h. Fluorescence was observed under a confocal microscope, and representative figures are shown. Quantification of RFP-LC3 and EGFP-2xFYVE positive aera are shown. n > 20 cells per group. All data are presented as mean ± SD. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ns, no significance; one-way ANOVA for multiple comparison and Tukey's test as post hoc test.