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. 2020 Dec 11;11(12):1047. doi: 10.1038/s41419-020-03251-w

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© The Author(s) 2020

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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Fig. 4 — Recombinant SEMG1 and SEMG2 bind PKM2, whereas SEMG1 only binds LDHA in GST pull-down assay followed by western-blotting. B 3×Flag-tagged SEMG1 and SEMG2 bind PKM2, whereas SEMG1 only binds LDHA in co-immunoprecipitation. C 3×Flag-tagged PKM2 interacts with both endogenous SEMG1 and SEMG2 in H520 cells, whereas 3xFlag-tagged LDHA binds SEMG1 only (co-immunoprecipitation). D Overexpression of SEMG1 and SEMG2 in H1299 cell line does not alters the mRNA levels of PKM2 and LDHA (Real-Time PCR). The stable overexpression of SEMG1 and SEMG2 increase the protein level and enzymatic activity of PKM2, whereas the overexpression of SEMG1 only elevates the protein level and enzymatic activity of LDHA in E, F H1299 cells and G, H Mia-Paca 2 cells. Knockdown of SEMG1 and SEMG2 in H520 cells decreases the protein level (I) and enzymatic activity (J) of PKM2 and LDHA. Three biological replicates were used for all quantifications, data are presented as mean ± S.D., *P < 0.05.