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. 2009 May 15;284(20):13843–13855. doi: 10.1074/jbc.M808515200

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Copyright © 2009, The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 8. — Autophagy is involved in mitochondrial morphology changes elicited by PINK1 deficiency and plays a neuroprotective role. A, stable vector control or PINK1shRNA cells co-transfected with the indicated siRNAs and quantified for the number of GFP-LC3 puncta per cell as analyzed in a representative experiment of two (, p < 0.001 versus control cell line; , p < 0.01 versus control siRNA; means ± S.E., n = 30-40 cells quantified per condition). Inset shows a representative Western blot showing effects of Atg7 or control siRNA. Note that knockdown of ATG7 decreases endogenous Atg7 levels and reduces the levels of activated LC3-II. B, the average integrated intensity of MitoSOX fluorescence was quantified in control or PINK1shRNA stable cell lines transiently transfected with the indicated siRNAs (, p < 0.01 versus control cell line (Ctrl), means ± S.E., n = 300-400 cells/condition). C and D, quantification of mitochondrial interconnectivity (C) and mitochondrial elongation (D) in PINK1 knockdown clonal lines transfected with the indicated siRNAs (representative of three experiments; , p < 0.05 versus control cell line transfected with control siRNA; , p < 0.01 versus PINK1 A4 or D14 transfected with control siRNA; n = 25-30 cells/condition). E, basal cell survival (Alamar Blue assay) of control and PINK1 shRNA clones (representative of four experiments; , p < 0.005 versus control stable cell line; n = 8 wells analyzed/condition). F, propidium iodide (PI) cell death assay of control and PINK1 shRNA stable clonal cell lines (, p < 0.05 versus control stable line; n = 800-1000 cells analyzed/condition). G, cell survival of control or PINK1 shRNA stable cell lines treated with bafilomycin or vehicle for 18 h (representative of three experiments; , p < 0.05 versus control cell line/bafilomycin; n = 4 wells analyzed/condition). H, cell survival of control or PINK1shRNA cells transfected with Atg7 siRNA (representative of three experiments; , p < 0.05 versus control siRNA-treated clone A4; , p < 0.005 versus control siRNA-treated clone D14; n = 6 wells analyzed/condition).

Inline graphic — Autophagy is involved in mitochondrial morphology changes elicited by PINK1 deficiency and plays a neuroprotective role. A, stable vector control or PINK1shRNA cells co-transfected with the indicated siRNAs and quantified for the number of GFP-LC3 puncta per cell as analyzed in a representative experiment of two (, p < 0.001 versus control cell line; , p < 0.01 versus control siRNA; means ± S.E., n = 30-40 cells quantified per condition). Inset shows a representative Western blot showing effects of Atg7 or control siRNA. Note that knockdown of ATG7 decreases endogenous Atg7 levels and reduces the levels of activated LC3-II. B, the average integrated intensity of MitoSOX fluorescence was quantified in control or PINK1shRNA stable cell lines transiently transfected with the indicated siRNAs (, p < 0.01 versus control cell line (Ctrl), means ± S.E., n = 300-400 cells/condition). C and D, quantification of mitochondrial interconnectivity (C) and mitochondrial elongation (D) in PINK1 knockdown clonal lines transfected with the indicated siRNAs (representative of three experiments; , p < 0.05 versus control cell line transfected with control siRNA; , p < 0.01 versus PINK1 A4 or D14 transfected with control siRNA; n = 25-30 cells/condition). E, basal cell survival (Alamar Blue assay) of control and PINK1 shRNA clones (representative of four experiments; , p < 0.005 versus control stable cell line; n = 8 wells analyzed/condition). F, propidium iodide (PI) cell death assay of control and PINK1 shRNA stable clonal cell lines (, p < 0.05 versus control stable line; n = 800-1000 cells analyzed/condition). G, cell survival of control or PINK1 shRNA stable cell lines treated with bafilomycin or vehicle for 18 h (representative of three experiments; , p < 0.05 versus control cell line/bafilomycin; n = 4 wells analyzed/condition). H, cell survival of control or PINK1shRNA cells transfected with Atg7 siRNA (representative of three experiments; , p < 0.05 versus control siRNA-treated clone A4; , p < 0.005 versus control siRNA-treated clone D14; n = 6 wells analyzed/condition).