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. 2021 Feb 1;9:620883. doi: 10.3389/fcell.2021.620883

Figure 5.

Figure 5

TLR2 is required for HMGB1-induced GSCs formation. (A) Expression of TLR2, TLR4, TLR9, RAGE in primary glioma cells after HMGB1 stimulation (800 ng/ml) was determined by qRT-PCR (n = 3). (B) Expression of TLR2 in primary glioma cells after HMGB1 stimulation (800 ng/ml) was determined by western blot (n = 3). (C) qRT-PCR analyses of the levels of CD133, OCT4, SOX2, and NANOG in TLR2 knockdown primary GBM cells treated with 800 ng/ml rhHMGB1 (n = 3). (D,E) Western blot analyses of the levels of CD133, OCT4, SOX2, and NANOG in TLR2 knockdown primary GBM cells treated with 800 ng/ml rhHMGB1 (n = 3). (F) Primary glioma cells were treated with HMGB1 (800 ng/ml) and transfected with siRNAs to TLR2, and then cultured under the neurosphere condition for 7 d. Number and diameter of tumor spheres on day 7 were quantified. Data are represented as mean ± SEM, *P < 0.05; **P < 0.01; ***P < 0.001.