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. 2017 Sep 22;7:12221. doi: 10.1038/s41598-017-12557-3

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© The Author(s) 2017

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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Identification of key genes associated with 5-aminolevulinic acid (5-ALA) fluorescence intensity in glioblastoma. (A) Results of RNA-sequencing of classified samples, which were analyzed according to differential expression along with the fluorescence intensity (fold change >2, p < 0.05, FDR < 0.01). Seventy-seven genes with positive correlation and 509 genes with negative correlation between the expression level and fluorescence intensity were identified. (B) Pathway analysis using gene set enrichment analysis (GSEA) resulted in 30 significantly enriched gene sets of (p < 0.01, FDR < 0.25). Among them, GLS2 (red arrows) was repeatedly found with a high enrichment score in the selected gene sets of metabolism and the neuronal system. (C) From the results of the RNA-seq FPKM values, GLS2 was highly expressed in areas of no fluorescence intensity, and the expression was decreased in areas of positive fluorescence in all samples. (D) The expression level of GLS2 in all samples, as measured by quantitative real-time PCR, confirmed the result of RNA-seq.