Fig. 3

The effects of tianeptine on the expression of autophagic proteins and autophagosome formation in B27-deprived hippocampal cells. Cells were treated with tianeptine (TIA; 50 and 100 µM) in the absence of growth medium B27 (B27 −) for 3 days. Western blots were employed to assess the levels of ULK phosphorylation (n = 8) (A), Beclin 1 (n = 10) (B), the LC3B-II/I ratio (n = 8) (C), and p62 (n = 8) (D). Quantitative analyses were normalized to the total ULK or the α-tubulin band. Full-length uncropped gels for all Western blots are available in the Supplementary Information. The cells were stained with the Cyto-ID Green autophagy dye and analyzed by confocal microscopy (n = 30 fields) (E). The values are expressed as a percentage of the control (B27 +/TIA −) and presented as the mean ± standard error of the mean (SEM). *p < 0.05, **p < 0.01 and ***p < 0.001 versus control; ^†p < 0.05, ^††p < 0.01 and ^†††p < 0.001 versus B27 deprivation only (B27−/TIA−).