Fig. 7.

HMGB1, another DAMP, promoted GSCs formation by primary glioma cells. a Primary glioma cells were cultured in the presence of CpG-ODN or HMGB1. The expression of stemness-related transcription factors including c-MYC, SOX2, NANOG, and OCT4 was determined by RT-qPCR, with β-actin as a reference control. The following concentrations were used: HMGB1, 1 μg/ml; CpG-ODN, 2 μM. b Primary glioma cells were cultured under the neurosphere condition in the presence of HMGB1 (1 μg/ml) for 7 days and photographed. The number of tumor spheres was counted. c Primary glioma cells were cultured in the presence of HMGB1 (1 μg/ml) and transfected with siRNAs toTLR9. Cells were then cultured for 3 days. The expression of stemness-related transcription factors including c-MYC, SOX2, NANOG, and OCT4 was determined by RT-qPCR, with β-actin as a reference control. d Primary glioma cells were cultured in the presence of HMGB1 and the expression of NEAT1 was determined by RT-qPCR, with β-actin as a reference control. Data are represented as mean ± SEM, n = 6. *, P < 0.05. **, P < 0.01. ***, P < 0.001. n.s, not significant