Fig. 3.

Effect of the autophagy agonist rapamycin on OGD/R-induced injury to SH-SY5Y cells. a Representative images showing the cell morphology of control cells, cells treated with 3 mM rapamycin (Rapa), and cells subjected to OGD/R with or without treatment using 3 mM Rapa 2 h before and during OGD/R. Scale bar is 100 μm. b Summary of the mean cell viability determined by MTT from three independent experiments using six wells of cells in each experiment. c Representative fluorescent images showing TUNEL staining (green), DAPI nuclear staining (blue), and merged images under the conditions shown in a. Scale bar is 100 μm. d Summary of the percentage of TUNEL-positive dead cells as shown in c, from three independent experiments with five different images analyzed in each experiment. e, f Analysis of cell death using flow cytometry under the conditions indicated above. g, h Western blotting showing the expression of LC-3 protein in SH-SY5Y cells. The mean results are from three independent experiments. *P < 0.05; **P < 0.01, and ***P < 0.001 vs. control group. ^##P < 0.01 and ^###P < 0.001 vs. OGD/R group