Fig 1: Elevated Dystroglycan Correlates with GBM Patient Outcome and aDG is Abundantly Glycosylated in GBM. aDAG1 expression was correlated with GBM patient survival using the Rembrandt (n = 523) and TCGA (n = 453) databases. b QPCR analysis of DAG1, ITGA6, EPHA2 and EPHA3 mRNA expression in GBM tissue specimens (n = 28). c Flow cytometric analysis for aDG glycosylation (IIH6 mAb) was performed on primary GBM cell lines grown as serum-free GNS cultures, compared to isotype control. See also Online Research 1d for full analysis. d aDG glycosylation was assessed by western blot in four primary GBM cell lines. e aDG glycosylation was assessed by western blot following cell fractionation to compare cytoplasmic versus membrane localisation. ß-actin was used as a loading control. f Flow cytometric analysis was performed for aDG glycosylation (IIH6 mAb) and EphA3 (IIIA4 mAb) in 10 early passage primary GBM cultures, mean channel fluorescence (mcf) was used to determine the correlation coefficient between EphA3 and glycosylated aDG (r = 0.899). GBM subtypes: MES mesenchymal, PN proneural, CL classical
Fig 2: DAG1 down regulation delays or prevents GBM formation in-vivo.a IF staining and QPCR was used to assess DAG1 mRNA levels following shRNA mediated KD in primary GBM cell lines. b Cell proliferation was assessed by direct cell counts using a haemocytometer following stable DAG1 KD. Data presented as the mean ± SD of three independent experiments, *p < 0.05. c Bright field images of WK1 and JK2 neurospheres 2 week post-DAG1 shRNA KD compared to control shRNA transfected cells. See also Online Resource 6a for SJH1 images. d Kaplan Meier analysis showing a significant p < 0.05 increase in overall survival following orthotopic intracranial engraftment of 1 × 105DAG1 shRNA versus control shRNA cells into the right striatum of NOD-SCID mice. WK1 n = 4 animals per arm, JK2 n = 7 animals per arm. e Representative H&E coronal sections from DAG1 shRNA versus control shRNA engrafted animal following euthanasia from either illness or tumour burden. See Online Resource 6d for WK1 H&E images
Supplier Page from Abcam for Anti-Alpha Dystroglycan antibody [2238]