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(A) Differentially expressed genes and overall expression trends of cortical neurons from seven different groups. “D vs A”: log₂FC of KCls vs Control, “G vs D”: log₂FC of MDZs vs KCls. (B) Gene expression profiles of commonly secreted neuronal factors under KCl, and MDZ conditions. <t>IGF1</t> is highlighted by a red dashed line. (C) Validation of IGF1and BDNF expression by RT-qPCR. n.s. P >0.05, * P < 0.05, ** P < 0.01, **** P < 0.0001. n=3. (D-E) Representative images and quantification of EdU assay after IGFBP3 experiment in GL261 cells. Scale bar = 100 μm. (F-G) Representative images and quantification of EdU assay after Linsitinib (OSI-906) treated experiments in GL261 cells. Scale bar = 200 μm. (H-I) Activation of the IGF1 downstream PI3K/AKT signaling pathway in glioma cells and its quantitative analysis. (J-K) PI3K/AKT signaling pathway change after IGFBP3 incubation in glioma cells. n.s. P >0.05, * P< 0.05, ** P< 0.01, *** P< 0.001, **** P< 0.0001. n=3.
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(A) Differentially expressed genes and overall expression trends of cortical neurons from seven different groups. “D vs A”: log₂FC of KCls vs Control, “G vs D”: log₂FC of MDZs vs KCls. (B) Gene expression profiles of commonly secreted neuronal factors under KCl, and MDZ conditions. IGF1 is highlighted by a red dashed line. (C) Validation of IGF1and BDNF expression by RT-qPCR. n.s. P >0.05, * P < 0.05, ** P < 0.01, **** P < 0.0001. n=3. (D-E) Representative images and quantification of EdU assay after IGFBP3 experiment in GL261 cells. Scale bar = 100 μm. (F-G) Representative images and quantification of EdU assay after Linsitinib (OSI-906) treated experiments in GL261 cells. Scale bar = 200 μm. (H-I) Activation of the IGF1 downstream PI3K/AKT signaling pathway in glioma cells and its quantitative analysis. (J-K) PI3K/AKT signaling pathway change after IGFBP3 incubation in glioma cells. n.s. P >0.05, * P< 0.05, ** P< 0.01, *** P< 0.001, **** P< 0.0001. n=3.

Journal: bioRxiv

Article Title: Midazolam suppresses glioma progression by attenuating neuronal activity and downregulating IGF1 signaling

doi: 10.64898/2026.03.31.715727

Figure Lengend Snippet: (A) Differentially expressed genes and overall expression trends of cortical neurons from seven different groups. “D vs A”: log₂FC of KCls vs Control, “G vs D”: log₂FC of MDZs vs KCls. (B) Gene expression profiles of commonly secreted neuronal factors under KCl, and MDZ conditions. IGF1 is highlighted by a red dashed line. (C) Validation of IGF1and BDNF expression by RT-qPCR. n.s. P >0.05, * P < 0.05, ** P < 0.01, **** P < 0.0001. n=3. (D-E) Representative images and quantification of EdU assay after IGFBP3 experiment in GL261 cells. Scale bar = 100 μm. (F-G) Representative images and quantification of EdU assay after Linsitinib (OSI-906) treated experiments in GL261 cells. Scale bar = 200 μm. (H-I) Activation of the IGF1 downstream PI3K/AKT signaling pathway in glioma cells and its quantitative analysis. (J-K) PI3K/AKT signaling pathway change after IGFBP3 incubation in glioma cells. n.s. P >0.05, * P< 0.05, ** P< 0.01, *** P< 0.001, **** P< 0.0001. n=3.

Article Snippet: The concentrations of IGF1 in Neu-CMs were quantified using a commercial Mouse/Rat IGF1 ELISA Kit (Multi Sciences, China) according to the manufacturer’s instructions.

Techniques: Expressing, Control, Gene Expression, Biomarker Discovery, Quantitative RT-PCR, EdU Assay, Activation Assay, Incubation

(A) Overall distribution of IGF1 - related genes in all samples. (B) Standardized expression profile and function annotation of IGF1co-expression genes. (C-D) Over-representation enrichment analysis of genes of interest using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases in KCl treated group and MDZ treated group, respectively. (E) Fuzzy cluster analysis identifying five distinct expression patterns of genes participated in MAPK signaling pathway. (F) Transcriptional activity of Fos was evaluated using a univariable linear model. (G) Predicted c-Fos binding motif within the Igf1 promoter region identified using the JASPAR database. (H) ChIP-qPCR analysis assessing c-Fos enrichment at the predicted regulatory region. * P <0.05, ** P < 0.01, **** P< 0.0001. n=3.

Journal: bioRxiv

Article Title: Midazolam suppresses glioma progression by attenuating neuronal activity and downregulating IGF1 signaling

doi: 10.64898/2026.03.31.715727

Figure Lengend Snippet: (A) Overall distribution of IGF1 - related genes in all samples. (B) Standardized expression profile and function annotation of IGF1co-expression genes. (C-D) Over-representation enrichment analysis of genes of interest using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases in KCl treated group and MDZ treated group, respectively. (E) Fuzzy cluster analysis identifying five distinct expression patterns of genes participated in MAPK signaling pathway. (F) Transcriptional activity of Fos was evaluated using a univariable linear model. (G) Predicted c-Fos binding motif within the Igf1 promoter region identified using the JASPAR database. (H) ChIP-qPCR analysis assessing c-Fos enrichment at the predicted regulatory region. * P <0.05, ** P < 0.01, **** P< 0.0001. n=3.

Article Snippet: The concentrations of IGF1 in Neu-CMs were quantified using a commercial Mouse/Rat IGF1 ELISA Kit (Multi Sciences, China) according to the manufacturer’s instructions.

Techniques: Expressing, Activity Assay, Binding Assay, ChIP-qPCR