Cytoplasmic expression of Hsp16.2 in different human brain tumors. Endogenous cytoplasmic expression levels of Hsp16.2 were assessed by Western blotting utilizing a custom made polyclonal anti-Hsp16.2 primary antibody. 30 samples were used including three from normal brain tissue and 18 samples from nine different types of brain cancer, two samples from each type. The subcellulare fractionation was confirmed by probing with antibodies recognizing nuclear H3 histone, cytoplasmic actin and equal loading was confirmed by a second incubation with anti-GAPDH antibody (data not shown). 1: normal brain, 2: Pilocytic astrocytoma (Grade 1), 3: Meningothelial meningioma (Grade1) 4: Diffuse astrocytoma (Grade 2), 5: Ependymoma (Grade 2), 6: Malignant meningioma (Grade 3), 7: Anaplastic astrocytoma (Grade 3), 8: Medulloblastoma (Grade 4), 9: Giant cell glioblastoma (Grade 4), 10: PNET (Grade 4).