Immunohistochemical co-staining for TLR2 and TLR9 with NeuN and GFAP. IHC co-staining of TLR2 and TLR9 with specific markers for neurons (NeuN) and astrocytes (GFAP) was performed and observed under a confocal microscope. The first antibodies used were rabbit anti-TLR2, rabbit anti-TLR9, rat anti-GFAP and mouse Fluor 488 conjugated anti-NeuN. The secondary antibodies used were Texas red conjugated donkey antibody to rabbit IgG and Cy2 conjugated goat antibody to rat IgG. The images show that TLR2 staining co-localized with neurons in the cortex of injured brain (A, B, C). However, co-localization of TLR2 with astrocytes was hardly observed in the brain section (D, E, F). Weak expression of TLR9 was observed co-localized with neurons in cortex of the injured brain (G, H, I). Co-localization of TLR9 with astrocytes was detected in the white matter of periventricular and subcortical regions of the brain (J, K, L).