Fig. S5.

Absence of mitochondrial abnormalities in mouse cortex overexpressing PARIS. (A) TEM images after stereotaxic injection of AAV1-GFP or (B) AAV1-PARIS into cortex. No apparent morphological change was found in between groups. N, nucleus (n = 3). (C) Western blots from total cell lysates of mouse cortical neurons transduced with lentiviral GFP or PARIS. Despite robust PARIS overexpression there is no change in the levels of SDHA, NDUFA9, or COXIV relative to β-actin. (D) TEM image analysis shows no alteration of mitochondrial number in AAV1-PARIS–injected cortex compared with AAV1-GFP–injected cortex (GFP; n = 95 cells, PARIS; n = 92 cells). The number of mitochondria harboring clear two-layer membranes and size larger than 0.15 μm² was counted. (E) Mitochondria in AAV1-PARIS–injected cortex are comparable to those in AAV1-GFP–injected cortex. The size of mitochondria in the cells randomly selected from cells with normal appearing nuclei (GFP; n = 352 mitochondria from 16 cells, PARIS; n = 321 from 14 cells) was measured by TEM software. The results were evaluated for statistical significance by applying the Mann–Whitney test (D and E). Data are expressed as median with interquartile range (75th and 25th quartiles).