Figure 5From: Intrathecal heat shock protein 60 mediates neurodegeneration and demyelination in the CNS through a TLR4- and MyD88-dependent pathway Intrathecal HSP60 does not result in major glial activation or proliferation in the cerebral cortex . Three days after intrathecal injection of 40 μg HSP60 or 40 μg SA (control) coronal sections of C67BL/6 J mouse brains were evaluated in terms of glial morphology and numbers. (A) Immunostaining of the cerebral cortex of C67BL/6 J mice (HSP60 n = 8, SA n = 5) with an Iba1 antibody. Scale bar, 100 μm, insets 50 μm. (B) Measurement of mean Iba1+ fluorescence intensity (HSP60 n = 4, SA n = 4). (C) Quantification of Iba1+ cells (HSP60 n = 8, SA n = 5). (B, C) Median, Mann–Whitney U test for the comparison of the HSP60-injected group with the SA-treated (control) group. (D) Immunostaining of the cerebral cortex of C67BL/6 J mice (HSP60 n = 7, SA n = 8) with a GFAP antibody. Scale bar, 100 μm, insets 50 μm. (E) Measurement of mean GFAP+ fluorescence intensity (HSP60 n = 4, SA n = 4). (F) Quantification of GFAP+ cells (HSP60 n = 7, SA n = 8). (E, F) Median, Mann–Whitney U test for the comparison of the HSP60-injected group with the SA-treated (control group). (G) Representative micrographs of the cerebral cortex immunostained with Iba1, GFAP, and Ki67 antibodies to mark microglia, astrocytes, and proliferating cells, respectively (HSP60 n = 4, SA n = 4). The subgranular zone of the hippocampal dentate gyrus served as a positive control for Ki67 reactivity. Scale bar, 100 μm.Back to article page