Skip to main content
. 2020 Dec 3;9(12):2585. doi: 10.3390/cells9122585

Figure 4.

Figure 4

Role of TLR4 on HMGB1 and CM-impaired function of glutamate clearance in primary astrocytes. (A) Effects of TLR4 knockdown on rHMGB1-reduced expression of GLAST. TLR4 and control siRNA-treated astrocytes were exposed to rHMGB1 (40 ng/mL) for 24 h and the expression levels of GLAST were examined using Western blot. (B) Effects of TLR4 inhibitors on rHMGB1-inhibited activity of astrocytic glutamate clearance. Primary astrocytes were exposed to rHMGB1 (40 ng/mL) with or without TLR4 inhibitors, eritoran or LPS-RS, and the activity of glutamate clearance was assayed 24 h later. (C) Primary astrocytes were exposed to CM with or without TLR4 inhibitors, eritoran or LPS-RS, for 24 h. Thereafter, the expression levels of GLAST protein were examined using Western blot. (D) Activity of astrocytic glutamate clearance was assayed by measuring the residual glutamate in CM. ## p < 0.01 vs. rHMGB1 alone, ** p < 0.01 vs. CM alone. Data are presented as mean ± SEM, n = 6. OGD/R, oxygen-glucose deprivation/reoxygenation; HMGB1, high-mobility group box 1; rHMGB1, recombinant HMGB1; CM, conditioned medium; TLR4, toll-like receptor 4; GLAST, glutamate aspartate transporter; n, number of repeated examinations; LPS-RS, lipopolysaccharide from Rhodobacter sphaeroides.