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. 2022 Feb 7;82(7):1283–1297. doi: 10.1158/0008-5472.CAN-21-2161

Figure 1.

Figure 1. LAMP2 is overexpressed in human GBM and GSCs. A, Representative immunoblot of LAMP2A, SOX2, SOX9, and β-actin in a set of glioma (U87, U373, U251, T98, A172) and patient-derived glioma stem (GNS166, GNS179, GNS144, GB2) cell lines (n = 4). B, Representative image of Western blot analysis for LAMP2A and β-actin in parental and 2ry oncospheres of glioma cell lines (n = 3). C, Representative images of the IHC staining of LAMP2A in subcutaneous tumors from parental U373 and respective oncospheres (n = 2). Scale bar, 100 μm. D, Single-cell RNA-seq results for the expression of LAMP2 among different cell types in glioblastoma samples (https://cells.ucsc.edu/?ds=gbm). E, LAMP2 mRNA expression in control and GBM samples in TCGA (P = 2.4e−13) and Rembrandt (P = 4.5E−03) cohorts. F, mRNA expression of LAMP2 in classical (cla), mesenchymal (me), and proneural (pro) subtypes of GBM from TCGA cohort (Pcla-me = 2.9e−09, Pcla-pro = 2.9e–08, Pme-pro = 4.5e−27) and Rembrandt cohort (Pcla-me = 4.8E−02, Pcla-pro = n.s., Pme-pro = 6.3E−04). G, LAMP2 mRNA expression in grade II–IV of glioma in TCGA (PII–III = n.s., PII–IV = 1.3e−02, PIII–IV = 7.5e−03) and Rembrandt (PII–III = n.s, PII–IV = 3.5e−05, PIII–IV = 3.1e−08) cohorts. H, LAMP2 expression in TMZ treatment of nonresponder (n = 154) and responder (n = 165) patients with GBM (http://www.rocplot.org/). Mann–Whitney test P value: 0.036. I, ROC curve of LAMP2 gene expression in patients with GBM treated with TMZ (n = 319; http://www.rocplot.org/). J, mRNA expression of LAMP2A, LAMP2B, and LAMP2C in GBM samples (n = 20) compared with a mix of healthy brain tissues. K, Representative tissue microarray and overall quantification for LAMP2A protein expression in GBM samples (n = 98). L and M, Representative image and quantification of CMA activity as the average number of puncta per cell (n > 1,500 cells per well in 4–6 independent wells) after transducing control (NIH3T3, N2A), GSCs (GNS166 and 179), and indicated GBM cell lines with photo switchable KFERQ-Dendra CMA reporter. Scale bar, 10 mm. Differences with NIH3T3 cells are compared. *, P ≤ 0.05; ***, P ≤ 0.001; ****, P < 0.0001.

LAMP2 is overexpressed in human GBM and GSCs. A, Representative immunoblot of LAMP2A, SOX2, SOX9, and β-actin in a set of glioma (U87, U373, U251, T98, A172) and patient-derived glioma stem (GNS166, GNS179, GNS144, GB2) cell lines (n = 4). B, Representative image of Western blot analysis for LAMP2A and β-actin in parental and 2ry oncospheres of glioma cell lines (n = 3). C, Representative images of the IHC staining of LAMP2A in subcutaneous tumors from parental U373 and respective oncospheres (n = 2). Scale bar, 100 μm. D, Single-cell RNA-seq results for the expression of LAMP2 among different cell types in glioblastoma samples (https://cells.ucsc.edu/?ds=gbm). E,LAMP2 mRNA expression in control and GBM samples in TCGA (P = 2.4e−13) and Rembrandt (P = 4.5E−03) cohorts. F, mRNA expression of LAMP2 in classical (cla), mesenchymal (me), and proneural (pro) subtypes of GBM from TCGA cohort (Pcla-me = 2.9e−09, Pcla-pro = 2.9e–08, Pme-pro = 4.5e−27) and Rembrandt cohort (Pcla-me = 4.8E−02, Pcla-pro = n.s., Pme-pro = 6.3E−04). G,LAMP2 mRNA expression in grade II–IV of glioma in TCGA (PII–III = n.s., PII–IV = 1.3e−02, PIII–IV = 7.5e−03) and Rembrandt (PII–III = n.s, PII–IV = 3.5e−05, PIII–IV = 3.1e−08) cohorts. H,LAMP2 expression in TMZ treatment of nonresponder (n = 154) and responder (n = 165) patients with GBM (http://www.rocplot.org/). Mann–Whitney test P value: 0.036. I, ROC curve of LAMP2 gene expression in patients with GBM treated with TMZ (n = 319; http://www.rocplot.org/). J, mRNA expression of LAMP2A, LAMP2B, and LAMP2C in GBM samples (n = 20) compared with a mix of healthy brain tissues. K, Representative tissue microarray and overall quantification for LAMP2A protein expression in GBM samples (n = 98). L and M, Representative image and quantification of CMA activity as the average number of puncta per cell (n > 1,500 cells per well in 4–6 independent wells) after transducing control (NIH3T3, N2A), GSCs (GNS166 and 179), and indicated GBM cell lines with photo switchable KFERQ-Dendra CMA reporter. Scale bar, 10 mm. Differences with NIH3T3 cells are compared. *, P ≤ 0.05; ***, P ≤ 0.001; ****, P < 0.0001.