Figure 4

NFκB upregulates human PINK1 gene expression. (A) NFκB increases the endogenous human PINK1 mRNA level. HEK293 cells were transfected with either the NFκB expression vector or empty vector pMTF. RT-PCRs were performed using either primers specific to the human PINK1 coding sequence or the human β-actin coding sequence. (B) NFκB increases the endogenous human PINK1 protein levels. HEK293 cells transfected with NFκB were harvested 48 h after transfection for protein detection. Cell lysates were run on 10% glycine gel and images were collected by Licor. A significant increase of endogenous PINK1 was observed. (C) The endogenous human PINK1 protein level was dramatically increased by NFκB in SH-SY5Y cells. SH-SY5Y cells were transfected with NFκB p65 plasmids or the control vector pMTF and then harvested for protein detections. The human PINK1 protein and its proteolysis products ∆1-PINK1 (55kD) and ∆2-PINK1 (45kD) were all increased. β-actin acted as the internal control. (D) LPS treatment facilitated PINK1 expression in SH-SY5Y cells. Cells were harvested after being treated for 16 h and then subjected to Western blot. The human β-actin level was served as a control. Quantification was performed by Image J software. Values indicate means ± SEM. n = 3, *p <0.01 by Student’s t-test.