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. 2020 Jul 9;14:210. doi: 10.3389/fncel.2020.00210

Figure 3.

Figure 3

(A,B) Western blotting analysis of relative protein contents in the hippocampus. (A) Western blot of protein contents of toll-like receptor 4 (TLR4), NF-κB p65, and TNF-α in the sham, SAH, and SAH + NaHS groups. (B) Bar graphs showing the relative optical density normalized to β-Actin as determined by ImageJ. (C,D) Expression and localization of TLR4, NF-κB p65, and TNF-α. (C) Representative micrographs of immunohistochemistry and quantification of mean density analysis by Image-Pro Plus 6.0 showing different levels of expression of TLR4 and TNF-α in the hippocampus, and NF-κB p65 in the cell nucleus in the hippocampus in the sham, SAH, and SAH + NaHS groups (scale bar = 20 μm in all images in panel C). (D) Representative micrographs of double immunofluorescence staining; overlapped images show that the cells expressing TNF-α (red) mostly colocalized with TLR4 (green). Quantification analysis showed that the density (cells/HPF) of cells staining positively for both TLR4 and TNF-α was much lower in the SAH + NaHS group than in the SAH group (scale bar = 20 μm in all images in panel D). Data are means ± SD. n = 8; *p < 0.05, **p < 0.01, ***p < 0.001 vs. sham group; #p < 0.05, ##p < 0.01 vs. SAH group.