Journal: Redox Biology
Article Title: A novel role of KEAP1/PGAM5 complex: ROS sensor for inducing mitophagy
doi: 10.1016/j.redox.2021.102186
Figure Lengend Snippet: PGAM5 interferes with PINK1 processing independently of its phosphatase activity. A. PGAM5 mutant and isoform lacking phosphatase activity induce Parkin translocation. PC6 cells were transfected with Parkin-EYFP and PGAM5 wt, PGAM5 F244D or short PGAM5 isoform (isoform 2, UniProt #Q96HS1-2). ****P < 0.0001, n = 6 dishes, 20 fields per dish, Welch's ANOVA followed by Dunnett's T3 multiple comparisons test. B. PGAM5 mutant lacking PARL/OMA1 cleaving site does not induce Parkin translocation. PC6 cells were transfected with Parkin-EYFP and PGAM5 or PGAM5 S24F. ****P < 0.0001, n = 6 dishes, 20 fields per dish, Welch's ANOVA followed by Dunnett's T3 multiple comparisons test. C. TOMM7 overexpression increases the fraction of full-length PGAM5 in HEK293 cells expressing PGAM5-flag and TOMM7-myc. D. TOMM7 overexpression protects partially against PGAM5 induced Parkin translocation. PC6 cells were transfected with Parkin-EYFP, PGAM5 wt or/and TOMM7. ****P < 0.0001, n = 9 dishes, 20 fields per dish, Welch's ANOVA followed by Dunnett's T3 multiple comparisons test. E. OMA1 and PARL protect partially against PGAM5 induced Parkin translocation. PC6 cells were transfected with Parkin-EYFP, PGAM5 wt and PARL or OMA1. ****P < 0.0001, n = 6 dishes, 20 fields per dish, One-way ANOVA followed by Sidak's multiple comparisons test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Article Snippet: PARL-FLAG , Addgene , Cat# 13639.
Techniques: Activity Assay, Mutagenesis, Translocation Assay, Transfection, Over Expression, Expressing