Double immunostaining of TLR
with transcription factor nuclear factor κB (NF- κB) after spinal cord ischemia reperfusion (I/R) injury. (A) Representative micrographs show the colocation of TLR4 (red) and NF-κB (green) at 12 h and 36 h after I/R injury. Arrows delineate co-localization. Scale bars are 100 μm. (B) Histogram for quantification of co-localized cells (cells with yellow signals). (C) Quantification of the TLR4 immunoreactivity is presented as average of three fluorescence intensity (FI) of three independent experiments. Double immunohistochemistry showed NF-κB had similar profile as TLR4 immunoreactivity after I/R injury. The rats pretreated with minocycline, TAK-242, PDTC were suggested significantly decreased TLR4 immunoreactivity after I/R and the number of TLR4- NF-κB-positive cells, whereas above effects synthetically increased in rats receiving LPS. All data are presented as mean ± SEM (n = 8 per group). **P < .01 compared to Sham group; ##P < .05 compared to I/R group.