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. 2021 Apr 26;9(1):e002091. doi: 10.1136/bmjdrc-2020-002091

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© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/.

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Glyoxal treatment leads to lipid vesicle accumulation independently of AMPK. The assays were performed on fibroblasts from three different donors. Control: untreated cells; glyoxal: 0.6 mM; AICAR: 1 mM; glyoxal+AICAR: 0.6 mM glyoxal+1 mM AICAR. (A) Western blot analysis of AMPKα and P-AMPKα and measurement of the P-AMPKα/AMPKα ratio. (B) BODIPY 493/503 fluorescence in green, anti-PLIN2 staining in red, and nuclear DAPI staining in blue (scale bare=20 µm). (C) The mean number of lipid vesicles per cell for each fibroblast donor. The LDs were counted randomly (≥50 cells were counted per condition). A t-test is indicated in (A). One-way ANOVA is indicated in (C). **p<0.01; ns=non-significant. The data represent means±SD. AICAR, 5-aminoimidazole-4-carboxamideribonucleotide; ANOVA, analysis of variance; DAPI, 4',6-diaminido-2-phenylindole; LD, lipid droplet.