Figure 4.

Protective autophagy was activated to alleviate Mn‐induced cell injury. After cells were treated with Mn and pretreated with 3‐MA. (A) The CCK‐8 assay was used to measure cell viability by using the microplate reader at 450 nm shown in a bar graph. (B) The LDH release measured using the microplate reader at 490 nm is shown in a bar graph. (C) The early apoptosis in neuron cell by flow cytometry (FCM) is shown in a bar graph. (D) The percentage of single positive populations (FITC +/PI –) in quadrant Q4 was regarded as the early apoptosis rate. (E) The fluorescence signal change in normal and treatment cells after transfection with ad‐mCherry‐GFP‐LC3B adenovirus. (F) The measurement of autophagic vacuoles after staining with MDC detected by FCM is shown in a bar graph. (G) The bands of beclin‐1, LC3, p62 and β‐actin expression levels measured by using Western blotting assay in SH‐SY5Y cells and the protein beclin‐1, LC3 and p62 after Western blotting experiments expressions are shown in a bar graph.*P < .05, **P < .01, compared to controls; ^# P < .05, ^## P < 01, compared to the 100μM Mn‐treated group; H, high dose, L, low dose