Fig. 7.

Melatonin (MT) inhibits environmental stress-triggered BNIP3-dependent mitophagy in placental trophoblasts. (A–D) Pregnant mice in Cd and MT + Cd groups received intraperitoneal injection of Cd on GD8. In MT and MT + Cd groups, pregnant mice were treated with MT from GD7 to GD15. Pregnant mice in Ctrl group were treated with saline. n = 10–12 from 10 to 12 different pregnant mice. (A) Representative immunofluorescent images of mouse placenta from Cd and MT + Cd groups. Arrows: co-localizations of TOM20 with LC3B. The Hoechst was used to tagged the nucleus. Scale bar: 20 μm. (B) Quantification for yellow dots per cell. (C) Representative immunoblots of BNIP3, LC3B-I/II, HSP60 and COX IV proteins in mouse placenta. (D) Quantification for BNIP3, LC3B-II, HSP60 and COX IV. (E–H) Human JEG-3 cells were pretreated using MT before Cd administration (n = 3 per group). (E) Representative immunoblots of BNIP3, LC3BI/II, HSP60 and COX IV proteins in mouse placenta. (F) Quantification for BNIP3, LC3BI/II, HSP60 and COX IV. (G) Quantification for yellow dots per cell. (H) Representative immunofluorescent images from Cd-treated cells with or without MT pretreatment. The Hoechst was used to tagged the nucleus. Scale bar: 20 μm. Data are expressed as the mean ± SEM. Multiple comparisons were performed using ANOVA. The post hoc test is executed adopting Bonferroni or Tamhane's T2 method following the result of homogeneity of variance test. *P < 0.05, **P < 0.01 versus PBS/NS group, ^#P < 0.05, ^##P < 0.01 versus Cd group. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)