Fig. 5.

p66Shc regulates formation of MAM and mitophagy. A-B Representative TEM images A and quantifications B of mitochondrial ultrastructural changes in NP and GDM placentae, mitochondria (in purple) in close contact with ER (in blue) in which distance ≤ 25 nm were considered as MAM sites. Scale bar, 0.5 μm; N, nucleus. **P < 0.01. C-D Representative TEM images C and quantifications D of mitochondria (in purple) in close contact with ER (in blue) in which distances ≤ 25 nm was considered as contacts in 5 mM and 30 mM glucose treated HTR8/SVneo cells. Scale bar, 200 nm. **P < 0.01. E JC-1-based flow cytometry assay images and quantifications of fluorescence intensity distribution of 5 mM or 30 mM glucose treated HTR8/SVneo cells. **P < 0.01. F-G Representative TEM images F and quantifications G of Mito-ER contacts distance in control and p66Shc-KD cells. Scale bar, 200 nm. ****P < 0.0001. H JC-1-based flow cytometry assay images and quantifications of fluorescence intensity distribution of control or p66Shc-KD cells. *P < 0.05. I-J Western blot images and quantifications of protein levels of Parkin and PINK in (I) NP and GDM placentae and J 5 mM and 30 mM glucose treated HTR8/SVneo cells. *P < 0.05; **P < 0.01; ****P < 0.0001. K Volcano plot showing the mRNA expression changes upon p66Shc knockdown. L Western blot images and quantifications of protein levels of Parkin and PINK in control and p66Shc-KD cells. *P < 0.05