Figure 6.

Protein kinase RNA‐like ER kinase signalling pathway participates in the initiation of protective autophagy. After cells were transfected with LV‐PERK shRNA and treated with Mn. (A) The fluorescence signal change in normal and treatment cells after transfection with ad‐mCherry‐GFP‐LC3B adenovirus. (B) The measurement of autophagic vacuoles after staining with MDC detected by FCM is shown in a bar graph. (C) 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 expressions are shown in a bar graph. (D) SH‐SY5Y cells were treatment with 100 μmol/L Mn for 6, 12 and 24 h, and the binding of ATF4 to the LC3 promoter by quantitative ChIP analysis is shown in line graph. (E) After cells were transfected with PERK shRNA and treated with Mn, the binding of ATF4 to the LC3 promoter by quantitative ChIP analysis is shown in a bar graph. (F) SH‐SY5Y cells were treated with 100 μmol/L Mn for 6, 12 and 24 h, and the expression levels of LC3 mRNA measured by RT‐qPCR assay are shown in a line graph. (G) The expression levels of LC3 mRNA measured by RT‐qPCR assay are shown in a bar graph. *P < .05, **P <.01, compared to controls; ^# P < .05, ^## P < .01, compared to the 100 μmol/L Mn‐treated group