Figure 3

HSP60 induces demyelination dependent on TLR4 and MyD88 in vivo . Three days after intrathecal injection of 40 μg HSP60 or 40 μg SA (control) coronal brain sections of C57BL/6 J (WT, HSP60 n = 4, SA n = 4), TLR4^−/− (HSP60 n = 4, SA n = 4), and MyD88^−/− (HSP60 n = 4, SA n = 4) mice were evaluated for the myelination status. (A) Immunostaining of the cerebral cortex of WT, TLR4^−/−, and MyD88^−/− mice with an antibody directed against APC. Scale bar, 50 μm. Cut-outs at higher power from the micrographs. Scale bar, 50 μm. (B) Quantification of APC+ cells in the cerebral cortex of WT, TLR4^−/−, and MyD88^−/− mice. Median; p values were determined for indicated groups by Mann–Whitney U test. (C) Immunostaining of the cerebral cortex of injected WT, TLR4^−/−, and MyD88^−/− mice with an antibody against MBP. Scale bar, 50 μm. Cut-outs at higher power from the micrographs. Scale bar, 50 μm. (D) Analysis of mean MBP+ fluorescence intensity in the cerebral cortex of WT, TLR4^−/−, and MyD88^−/− mice. Median, Mann–Whitney U test for indicated groups.