Fig. 5
Loss of host-derived GPNMB impairs tumor growth in OBS cultures injected with glioma cells with low and high intrinsic expression of GPNMB. A RT-qPCR of GPNMB expression relative to cultured murine astrocytes (n = 3) in cultured RCAS-PDGFb cells (n = 3) and the GL261 glioma cell line (n = 3). Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparisons test. B Western Blot of total GPNMB protein isolated from cultured RCAS-PDGFb, GL261 glioma cells, cultured MG (neonatal) and MG (adult). GAPDH expression serves as a reference. Each column represents an individual batch of culture (n = 3). C OBS from WT and KO animals injected with cultured PDGFb-driven RCAS-RFP glioma cells. Images are shown on the left. Top row: Orthogonal view on RFP immunofluorescence based on z-stack and tile scanning. Bottom row: representative 3D-reconstruction in Imaris based on z-stack and tile scanning. The graph on the right summarizes the volumes (in mm3) of WT (n = 11) and KO (n = 9) tumors as determined from the scans. D OBS injected with cultured mCherry-GL261 glioma cells. Images are shown on the left. Top row: Orthogonal view on mCherry immunofluorescence based on z-stack and tile scanning. Bottom row: representative 3D-reconstruction in Imaris based on z-stack and tile scanning. The graph on the right summarizes the volumes (in mm3) of WT (n = 30) and KO (n = 42) tumors as determined from the scans. The scale bars represent 500 µm. Statistical analysis was performed using unpaired t-test. Error bars represent SD. ****p < 0.0001, ns, not significant