Fig. 6

TNF-α facilitates MB tumor progression in vivo. a Primary astrocytes from the normal cerebella of WT mice (pAS) and TAAs were isolated and prepared for TNF-α mRNA evaluation by qPCR, **p < 0.01. b Normal cerebella of WT mice (WT), primary MB tissue and subcutaneous MB tissue treated with the C3aR antagonist SB290157 or MCT control were harvested and homogenized in lysis buffer, and the tissue homogenate samples were centrifuged to collect the supernatant. Then, TNF-α protein levels in these supernatants were assessed by ELISA. ***p < 0.001 vs. WT group; ##p < 0.01 vs. MCT group. c Overall survival rate of MB patients in TNF-αR high (blue line) and low (red line) expression groups was followed up to 60 months. Analysis was performed using the R2 Genomics Analysis and Visualization Platform. d–g Subcutaneous MB-bearing mice were treated with the TNF-α antagonist R-7050 (30 mg/kg per mouse per day) or vehicle control (MCT) daily by intraperitoneal injection. Subcutaneous tumor volume was monitored every day, and the volume was curved based on the fold changes (d). At the termination of treatment, tumors were dissociated and photographed to show tumor size (e). Frozen sections were prepared from tumor tissues after treatment. Ki67 was immunostained red, and DAPI was used to counterstain the cell nuclei (f). The percentage of Ki67 +  cells in tumor sections was quantified (g), **p < 0.01. The results shown are representative of two independent experiments. Values are expressed as the mean ± SEM (n = 4 mice/group)