Fig. 1: Lysosomal activity is associated with the malignant phenotype of GBM. | Nature Communications

Fig. 1: Lysosomal activity is associated with the malignant phenotype of GBM.

From: Lysine-arginine imbalance overcomes therapeutic tolerance governed by the transcription factor E3-lysosome axis in glioblastoma

Fig. 1

A Detection of lysosomal proteolytic activity at the basal level with DQ-BSA in patient-derived GBM cell lines with confocal fluorescence microscopy (n = 30 cells/group). NPC, human neural progenitor cells. B The correlation analysis of the average sphere size formed in a medium containing 1% methylcellulose for 10 days and lysosomal proteolytic activity from A in GBM cell lines. Red: Lyso-high; Black: Lyso-low GBM cell lines (n = 3 independent experiments). C Detection of cathepsin B activity at the basal level in Lyso-high (red line) and Lyso-low (black line) GBM cell lines using flow cytometry (n = 3 independent experiments). D Measurement of lysosomal pH at the basal level in Lyso-high (red) and Lyso-low (black) GBM cell lines using flow cytometry (n = 3 independent experiments). E The representative images and quantification of galectin-3 punctation in GBM cells treated with 50 μM LLOMe for 15 min (n = 3 independent experiments). Scale bars, 10 μm. F GSEA results of RNA-seq data in the Lyso-high versus Lyso-low GBM cells. FDR < 25%. G, H Mitochondrial membrane potential (G) and mitochondrial ROS (H) at the basal level were detected with JC-10 and MitoSOX, respectively, and evaluated in Lyso-high (red line) and Lyso-low (black line) GBM cell lines (n = 3 independent experiments). I Representative bioluminescence images of mice bearing GBM cells after 6 and 27 weeks after transplantation (n = 3 mice/GBM cell line). J Kaplan–Meier survival curves for nude mice transplanted with Lyso-high (red) or Lyso-low (black) GBM cells (n = 3 mice/GBM cell line). Data are presented as the means ± SD. Statistical analysis (A, C, G, and H) using unpaired two-tailed Student’s t-tests. Statistical analysis (B) using Pearson correlation analysis. Statistical analysis (D) using two-tailed nested t-test. Statistical analysis (E) using one-way ANOVA. Statistical analysis (J) using Kaplan–Meier method with the log-rank test. ****p < 0.0001. Experiments were repeated at least three times with similar results. Source data are provided with this paper.

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