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. 2024 Sep 15;81(1):404. doi: 10.1007/s00018-024-05427-5

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 5 — c-Myc lactylation mediates the HSPA12A-increased c-Myc nuclear localization in TEC following H/R. A Scheme illustrates the process of c-Myc lactylation. p300 inhibitor C646 was used to inhibit c-Myc lactylation. B c-Myc lactylation. The anti–c-Myc immunoprecipitates were prepared from H/R-treated TEC followed by immunoblotted for L-lactyl-lysine (Klac). The IgG immunoprecipitates served as negative controls. Data are mean ± SD, **P < 0.01 by Student’s two-tailed unpaired t- test. n = 3. C Nuclear localization of lactylated and total c-Myc. Immunostaining for lactylation (Klac, green) and c-Myc (red) was performed in TEC after H/R. DAPI was used to stain nuclei (blue). Data are mean ± SD, **P < 0.01 by Student’s two-tailed unpaired t- test. Scale bar = 2 μm. n = 3. D Overlap efficiency of Klac and c-Myc in nuclei. This overlap efficiency was quantified from the images of (B). Data are mean ± SD, **P < 0.01 by Student’s two-tailed unpaired t- test. n = 3. TEC were challenged with H/R in the presence or absence of C646. The following experiments were performed subsequently. E c-Myc lactylation. Anti–c-Myc immunoprecipitates prepared from H/R-treated TEC were immunoblotted for lactylation antibody (Klac). Data are mean ± SD, **P < 0.01 by one-way ANOVA followed by Brown-Forsythe and Welch test. n = 6. F Nuclear localization of lactylated and total c-Myc. Immunostaining for lactylation (Klac, green) and c-Myc (red) was performed in H/R-treated TEC. DAPI was used to stain nuclei (blue). Data are mean ± SD, ** < 0.01 by one-way ANOVA followed by Tukey’s test. Scale bar = 2 μm. n = 3. Arrows showed reduced nuclear but increased perinuclear localization of c-Myc following inhibition of c-Myc lactylation. G mRNA levels. The mRNA expression of indicated genes were analyzed by RT-PCR. *P < 0.05 and **P < 0. 01 by one-way ANOVA followed by Tukey’s test. n = 4 for Rcc1 groups and n = 5 for all the other groups. H TEC proliferation. Cell proliferation was indicated by EdU incorporation following H/R. Data are mean ± SD, **P < 0.01 by one-way ANOVA followed by Tukey’s test. n = 3. Scale bar = 50 μm