Figure 7

HSP60 is predominantly expressed in CNS neurons and is upregulated after injury. (A) Immunofluorescence photomicrographs of the infarct (ipsilateral; ipsi and contralateral; co) and lesion-associated region (lar) in brains of C57BL/6 J mice at 1 d after MCAo. Sham-operated animals served as a negative control. Tissues were double-stained with anti-Iba1, anti-GFAP, anti-APC and anti-NeuN antibodies to mark microglia, astrocytes, oligodendrocytes, and neurons, respectively, and with anti-HSP60. Nuclei were stained with DAPI. Scale bar, 100 μm. Cut-outs at higher power from the micrographs. Scale bar, 10 μm. (B) Ipsi- and contralateral hemispheres of C57BL/6 J mice at 1 d after MCAo and of the respective sham-operated animals were analyzed by SDS-PAGE and subsequent immunoblotting with an HSP60 antibody. β-actin served as a housekeeping control. (C) Lysates of cultured neurons and microglia, both incubated with 10 μg/ml imiquimod or PBS (control) for 12 h and 3 h, respectively, and lysates of Oli-neu cells incubated with 10 μg/ml imiquimod, 1 μM staurosporine (St), or PBS (control) for 3 h were analyzed by SDS-PAGE and subsequent immunoblotting with an HSP60 antibody. β-actin served as a housekeeping control.