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. 2019 Jun 17;9:8671. doi: 10.1038/s41598-019-45191-2

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The senescent phenotype is associated with a switch to a glycolytic metabolism and mitochondria modifications. (A) ASC and dermal fibroblasts were treated (CAP or NT) and glucose and lactate concentrations were measured in supernatant 1 (D1) and 7 (D7) days after the treatment (n = 3 to 6). (B) The level of GLUT1, HKII, LDHA and MCT4 mRNA expression was quantified by RT-qPCR 7 days after the treatment (ASC n = 6; fibroblasts n = 3). (C) ASC were treated (CAP or NT) and tested after 1 and 7 days for their mitochondrial membrane potential using the TMRM probe, the mitochondria mass using the MitoTracker probe, intracellular ROS production using H₂DCFD and mitochondrial ROS production using Mitosox probe. Quantification is represented by the Mean of Fluorescence Intensity (MFI) normalized to the same condition without labelling (n = 4). (D) ASC were treated (CAP or NT) and tested for mitochondrial DNA quantification 1 and 7 days after treatment (n = 3). (E) ASC were treated (CAP or NT) and stained by immunofluorescence for mitochondria content 1 and 7 days after treatment. Pictures were taken with a Nikon, Eclipse, TE2000-S microscope (x400 magnification) with the same calibration and a zoom was realized with a confocal LSM 780 (Zeiss) (x630 magnification). Quantification is performed using the high content imaging system Operetta (Perkin Elmer, lens x20) (n = 3). Data represent mean ± SEM of independent experiments as indicated (n).