Fig. 3

In vitro LPS stimulation alters the immuno-phenotype of glioma cells and GSCs dependent on TLR4. a: TLR4 expression was stably knocked down in RG2 and U87 GSCs by specific shRNA as determined by western blot analyses. b-c: In Control-shRNA cells LPS stimulation for 6 h increased MHC-I, MHC-II, CD80, CD86, TNF-α, and IL-6 expression, but decreased IL-10 expression as determined by real time-PCR (b) and western blot (c). However, in the presence of TLR4-shRNA, the effects of LPS stimulation were abrogated. d: After LPS stimulation for 6 h, the levels of TNF-α, IL-6 and IL-10 secretion were detected by ELISA in cell-free supernatants from RG2-control-shRNA and RG2-TLR4-shRNA GSCs. e: CXCL10 was upregulated in RG2 GSCs after LPS stimulation for 6 h in a TLR4-dependent manner, as determined by immunofluorescence stain and western blot. Scale bars, 25 μm. Representative images are shown. Results are presented as the mean ± SEM of triplicate samples from three independent experiments. *P <0.05, **P <0.01, ***P <0.001