egfp rab10  (Addgene inc)

Journal: Science advances

Article Title: Retromer promotes the lysosomal turnover of mtDNA.

doi: 10.1126/sciadv.adr6415

Figure Lengend Snippet: Fig. 6. The small GTPase RAB10 promotes mitochondrial fragmentation and mtDNA degradation in lysosomes. (A) Immunostaining of HeLa cells expressing the constitutive active protein RAB10Q68L-GFP labeled with α-VPS35. (B) Manders’ correlation coefficient between RAB10 and VPS35. (C and D) Confocal images of cells ex- pressing WT RAB10-GFP, constitutive active RAB10Q68L-GFP, dominant negative RAB10T23N-GFP, in the steady state, and (D) expressing TWNKK319E-mCherry, labeled with α-TOM20. (E) Quantification of the mitochondrial morphology in RAB10 expressing cells (n = 3, >20 cells per replicate). (F and G) Cells expressing RAB10Q68L-GFP and (G) TWNKK319E-mCherry labeled with α-LAMP1 and α-dsDNA. Arrows depict RAB10-LAMP1-dsDNA foci. (H) Manders’ correlation coefficient between RAB10-GFP and LAMP1 and LAMP1 and dsDNA (n = 3, 10 images per replicate). (I) RAB10-GFP coimmunoprecipitation in the steady state and cells expressing TWNKK319E-mCherry with the lysosomal protein LAMP1. P values were calculated using one-way ANOVA with Tukey correction for multiple comparisons. Scale bars, 10 μm. Data are presented as means ± SEM.

Article Snippet: RAB10 plasmids were obtained from Addgene (WT, Addgene #49472; Q68L Addgene #49544; and T23N Addgene #130885).

Techniques: Immunostaining, Expressing, Labeling, Dominant Negative Mutation

Journal: Science advances

Article Title: Retromer promotes the lysosomal turnover of mtDNA.

doi: 10.1126/sciadv.adr6415

Figure Lengend Snippet: Fig. 9. Proposed model for retromer function upon mtDNA stress. The retromer enhances mitochondrial fragmentation and mtDNA turnover. mtDNA ejection occurs in a BAX-dependent manner, targeting RAB10-VPS35-positive lysosomes, and independent of MDVs. Stimulation of these pathways restores mitochondrial function and mitigates defects associated with mtDNA damage in vivo.

Article Snippet: RAB10 plasmids were obtained from Addgene (WT, Addgene #49472; Q68L Addgene #49544; and T23N Addgene #130885).

Techniques: In Vivo