Fig 2: NOK senescence displays important distinguishing characteristics. (A) Mass spectrometry analysis of proposed SASP additions. Selected proteins significantly increased in EV pellet or CM (Mean ± SD). Y-axis scales differ. (B) mRNA levels of proposed SASP additions. (Mean ± SD). Y-axis scales differ. (C) TEM of vesicles from EV pellet labelling for surface HSP60 (left, and 4x enlargement, right). For Figure 6A, 6B, *p<0.05, **p<0.01, ***p<0.001, **** p<0.0001.

Fig 3: Gestational exposure to environmental stress activates mitophagy in placental trophoblasts. (A–F) Each pregnant mouse in Cd group received intraperitoneal injection of Cd on GD8. Each pregnant mouse in Ctrl group was treated with saline. n = 10–12 from 10 to 12 different pregnant mice. (A) Representative immunoblots of HSP60, COX IV and PGC1a proteins in placentae. (B–D) Quantification for HSP60, COX IV and PGC1a. (E) Representative immunofluorescent images of mouse placentas from NS and Cd groups. Arrows: co-localizations of TOM20 with LC3B. The Hoechst was used to tagged the nucleus. Scale bar: 20 µm. (F) Quantification for yellow dots per cell. (G–J) Human JEG-3 cells were incubated with Cd for 0, 2, 6 or 12 h (n = 3 per group). (G) Representative immunoblots of HSP60, COX IV and PGC1a proteins in the cells. (H–J) Quantification for HSP60, COX IV and PGC1a. (K–M) Human JEG-3 cells were pretreated using CQ before Cd stimulation (n = 3 per group). (K) Representative immunoblots of HSP60 and COX IV proteins in the cells. (L and M) Quantification for HSP60 and COX IV. 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 < 0.01 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.)

Fig 4: EHMT2 controls the expression and subcellular localization of HSPD1. (A) Phosphor array with the lysates of cells in which EHMT2 was knocked down. The phosphor array (ARY003B) was purchased from R&D Biosystems. Approximately 200 µg of cell lysate was used. (B and C) Expression levels of HSPD1 in breast cancer samples (TCGA); P-values were calculated using Wilcoxon’s tests (***P<0.001) (B) and RNA-seq results (C). (D) RT-qPCR analysis of HSPD1 after EHMT2 knockdown in MDA-MB-231 cell lines. ACTB was used as the internal control (top panel). Western blot analysis was performed using the indicated antibodies (bottom panel). (E) Cell growth assay. After knocking down HSPD1 for 72 h, the cells were fixed with 100% methanol and stained with crystal violet. Scale bar, 200 µm (top panel); P-values were calculated using Student’s t-tests (**P<0.01) (bottom panel). (F) Western blot analysis after HSPD1 knockdown using anti-HSPD1, anti-PARP, anti-caspase3 and anti-ß-catenin antibodies. ACTB was used as the internal control in MB231 cells. (G) Immunocytochemical analysis of HSPD1. MB231 cells treated with siCont or siEHMT2 were fixed with 100% methanol and stained with anti-HSPD1 (Alexa Fluor 488, green) and DAPI (blue). Scale bar, 200 µm. (H) Cell growth assay. After MB231 cells were treated with BIX01294 for 24 h, the cells were fixed with 100% methanol and stained with crystal violet. Scale bar, 200 µm (top panel); P-values were calculated using Student’s t-tests (**P<0.01). (I) RT-qPCR analysis of HSPD1 after treatment with BIX01294. ACTB was used as the internal control (top panel), and the signal intensities corresponding to HSPD1 were quantified using ImageJ software (bottom panel). The P-values were calculated using Student’s t-test (**P<0.01). (J) Western blot analysis after treatment with BIX01294 using anti-HSPD1, anti-PARP, anti-caspase3 and anti-ß-catenin antibodies. ACTB was used as the internal control in MB231 cells.

Fig 5: ZWT promoted mitophagy in podocytes. (A, B) The protein blots and quantitative analysis of LC3 II/I, p62, HSP60, and TOMM20. The obtained values of HSP60, TOMM20, and p62 were normalized to GAPDH; LC3 II values were normalized to LC3 I. (C, D) The representative images and quantitative analysis of LC3 and COX IV colocalization (×400, scale bar = 20 µm). Data are represented as mean ± SD (n = 3). # p < 0.05, ## p < 0.01 versus control group, *p < 0.05 and **p < 0.01 versus TNF-a group.