Time of feeding affects daily protein O-GlcNAcylation rhythm in mouse liver.
<p><b>(A)</b> Schematics showing the study design. An 8-week-old C57BL/6 male mice were subjected to either NRF or DRF. The NRF group was only fed during the dark phase (ZT12–24), while the DRF group was only fed during the light phase (ZT0–12). After 3 weeks of treatment, mouse li...
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2025
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| Summary: | <p><b>(A)</b> Schematics showing the study design. An 8-week-old C57BL/6 male mice were subjected to either NRF or DRF. The NRF group was only fed during the dark phase (ZT12–24), while the DRF group was only fed during the light phase (ZT0–12). After 3 weeks of treatment, mouse livers were collected at 4 h intervals over one day, and global O-GlcNAcylation of liver nuclear proteins were detected. <b>(B)</b> Representative blot showing the daily rhythm of nuclear protein O-GlcNAcylation in liver samples from the NRF vs. DRF mice (Top panel). Total nuclear proteins were stained with Coomassie blue (Bottom panel) and used for normalization. <b>(C)</b> Quantification of protein O-GlcNAcylation signals in <b>(B)</b> (<i>n</i> = 3). The α-streptavidin signal of the entire lane was quantified. The NRF data is replotted from <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003400#pbio.3003400.g001" target="_blank">Fig 1B</a>. <b>(D)</b> Venn diagram to compare the O-GlcNAc sites with day vs. night difference (>20% fold change) in NRF and DRF liver. <b>(E–F)</b> Heatmaps showing the relative abundance of site-specific O-GlcNAcylation between ZT11 (day) and ZT23 (night). Heatmap is generated using detected O-GlcNAcylated peptides in <b>(E)</b> and O-GlcNAc sites in <b>(F)</b>. <b>(G)</b> Daily rhythm of UDP-GlcNAc were quantified using targeted metabolomics (<i>n</i> = 3). The NRF data is replotted from <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003400#pbio.3003400.g001" target="_blank">Fig 1C</a>. The dashed lines represent the median levels of O-GlcNAcylation <b>(C)</b> or UDP-GlcNAc <b>(G)</b> in NRF and DRF groups, and Student <i>t</i> test was used to assess significant difference. The gray shading illustrates the dark phase of a day–night cycle. Data are presented as mean ± SEM, and the shading area around the curves represents SEM. <b>(H)</b> Food consumption of NRF and DRF mice over a 24-h period (<i>n</i> = 5). <i>p</i> = 0.0952, Mann–Whitney test. The numerical values of all replicates in <b>(C)</b>, <b>(G),</b> and <b>(H)</b> can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003400#pbio.3003400.s023" target="_blank">S1 Data</a>.</p> |
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