quantitative elisa mouse rat igf1 kit liver glycogen assay kit  (R&D Systems)

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: After 24 h incubation in Neu-CMs with or without 1ug/ml IGF1 binding protein-3 (Sigma, USA) [ ] , 100 μM IGF 1R inhibitor Linsitinib (OSI-906) (Sellect, USA) [ , ] or exogenous IGF1 factor (MCE, USA), the proliferation of glioma cells was detected by EdU assay.

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

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: After 24 h incubation in Neu-CMs with or without 1ug/ml IGF1 binding protein-3 (Sigma, USA) [ ] , 100 μM IGF 1R inhibitor Linsitinib (OSI-906) (Sellect, USA) [ , ] or exogenous IGF1 factor (MCE, USA), the proliferation of glioma cells was detected by EdU assay.

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

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

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

Journal: Development (Cambridge, England)

Article Title: Kupffer cells control neonatal hepatic metabolism via Igf1 signaling

doi: 10.1242/dev.204962

Figure Lengend Snippet: KC-hepatocyte crosstalk is altered in P0 KO Spi1 livers. (A) Bar plot showing the relative information flow of between WT Spi1 and KO Spi1 of inferred cell–cell communication using CellChat. (B) Comparison of the significant ligand–receptor pairs between WT Spi1 and KO Spi1 , which contribute to the signaling from KCs to the hepatocyte clusters. (C) Heatmap showing the relative importance of KC and hepatocyte clusters as sender, receiver, mediator and influencer, based on the computed four network centrality measures of IGF (top) and visfatin (bottom) signaling. (D) Box plot of variance stabilizing transformation-normalized Igf1 expression in hepatocytes and macrophages in WT Spi1 and KO Spi1 mice at P0. n =5 per genotype from 3 independent litters. Differential expression was tested using DESeq2 on raw counts. The whiskers represent the 5-95% percentile, the box extends from the 25th to 75th percentiles and the horizontal line represents the median. (E) Serum insulin levels measured by ELISA on WT Spi1 and KO Spi1 at P0. n =4-5 per genotype from 4 independent litters. Bar plot presented as mean±s.d. Unpaired Student's t -test. (F) Serum glucagon levels measured by ELISA on WT Spi1 and KO Spi1 at P0. n =6 per genotype from 3 independent litters. Bar plot presented as mean±s.d. Unpaired Student's t -test. (G) Enrichment analysis of downregulated phosphorylation sites showing the decreased and increased phosphorylation in KO Spi1 liver compared to WT Spi1 . n =4-6 per genotype from 5 independent litters.

Article Snippet: To determine the amount of Igf1 present in the perinatal liver and serum the Quantitative ELISA Mouse/Rat Igf1 Kit Liver Glycogen Assay Kit from R&D Systems (MG100) was used.

Techniques: Comparison, Transformation Assay, Expressing, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay, Phospho-proteomics

Journal: Development (Cambridge, England)

Article Title: Kupffer cells control neonatal hepatic metabolism via Igf1 signaling

doi: 10.1242/dev.204962

Figure Lengend Snippet: KC-derived Igf1 regulates glycogen homeostasis in hepatocytes at birth. (A) Percentage of (left) and normalized (right) Igf1 expression in the respective hepatic cell type during embryogenesis. (B) Breeding scheme to produce KO Igf1 mice and littermate controls ( WT Igf1 ). Created in BioRender by Mass, E., 2025. https://BioRender.com/jvsfc8p . This figure was sublicensed under CC-BY 4.0 terms. (C,D) Igf1 levels measured by ELISA on whole liver lysate (C) and serum (D) of WT Igf1 and KO Igf1 at P0. n =7-8 per genotype from 4 independent litters. Bar plots presented as mean±s.d. Unpaired Student's t -test. (E) Glycogen levels measured on whole liver lysates of WT Igf1 and KO Igf1 at P0. n =11-16 per genotype from 7 independent litters. Values were normalized per litter. The whiskers represent the 5-95% percentile, the box extends from the 25th to 75th percentiles and the horizontal line represents the median. Cross indicates the mean. Mann–Whitney test. (F) Representative transmission electron micrograph from WT Igf1 and KO Igf1 livers at P0. n =3-4 per genotype from 2 independent litters. GP, glycogen particle; N, nucleus. Scale bars: 8 µm. (G) Scheme indicating the quantification process of glycogen content in hepatocytes. (H) Hepatocyte glycogen content of KO Igf1 normalized to WT Igf1 littermates; each value represents one hepatocyte (ten hepatocytes were assessed per liver). n =3-4 per genotype from 2 independent litters. Mann–Whitney test. (I) Normalized total metabolite abundance in WT Igf1 and KO Igf1 livers following [U- 13 C 6 ]-glucose tracing at P0. n =5-6 per genotype from 2 independent litters. Unpaired Student's t -test. ns, not significant ( P >0.05). (J) Fractional enrichment of labeled metabolites following [U- 13 C 6 ]-glucose tracing at P0 with and without the addition of exogenous Igf1 protein. Liver samples with and without Igf1 from the same animal are connected with a line. n =5-6 per genotype from 2 independent litters. Wilcoxon test.

Article Snippet: To determine the amount of Igf1 present in the perinatal liver and serum the Quantitative ELISA Mouse/Rat Igf1 Kit Liver Glycogen Assay Kit from R&D Systems (MG100) was used.

Techniques: Derivative Assay, Expressing, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Transmission Assay, Labeling