Figure 2. .

Immunohistochemical and immunofluorescent staining of normal human brain and muscle sections with ACAD9 antisera and human lung sections with LCAD antisera. a and b, Immunohistochemical staining of human muscle with VLCAD (a) and ACAD9 (b) antibody visualized with DAB (brown), counterstained with a blue nuclear stain. c, Diffuse and punctate pattern of ACAD9 staining, extending to neuronal dendrites of Purkinje cells. The nucleus of Purkinje and granular neurons does not stain with ACAD9 antibodies visualized with fluorescent secondary antibody (white). d–i, Normal human brain section immunostained with ACAD9 antibody and visualized with DAB (brown). All sections are counterstained with a nuclear stain (blue). d, Neurons of human neocortex. e, Neurons in the CA1 region of normal human hippocampus showing relatively weak ACAD9 staining. f, Neurons in the CA4 region and granular neurons of hippocampus. g, Cerebellar cortex, including Purkinje neurons, their dendrites, and neurons in the granular layer, showing relatively strong ACAD9 staining. h, Absence of ACAD9 staining in cerebellar white matter. i, Strong ACAD9 staining in dentate nucleus in the deep white matter of cerebellum. j and k, Normal human lung sections stained with LCAD (j) or the marker protein MUC1 (k) (a membrane protein, specific to human alveolar septae) antibody and visualized by fluorescent secondary antibodies, LCAD (j, red) and MUC1 ( k, green), colocalizing to type II alveolar epithelial cells (white arrow), as described in the “Methods” section. l and m, LCAD antibodies specifically staining bronchial epithelium and the alveolar septae of normal human lung tissue (l). At high magnification, the staining of LCAD in bronchial epithelium (m) shows a diffused punctate pattern consistent with mitochondrial staining.