Research Paper Volume 13, Issue 9 pp 13087—13107

Figure 1. Localization of mimecan expression in mouse adrenal glands using in situ hybridization and immunohistochemistry. (A) Positive staining (red) of sections of adrenal glands hybridized with DIG-labeled mimecan antisense probes primarily appeared in the adrenal cortex and some of the medullary mesenchyme (image obtained under visible light at 10× magnification). (B) The field shown is the same as (A) under fluorescent light. (C) Sections of adrenal glands hybridized with fluorescein-labeled mimecan and DIG-labeled PNMT antisense probes, confirming mimecan expression (red) primarily within the adrenal cortex and PNMT (phenylethanolamine-N methyl transferase; blue) expression in the medulla; no co-expression was observed (image obtained under visible light at 10× magnification). (D) The field shown is the same as (C) under fluorescent light. (E) Sections of adrenal glands hybridized with fluorescein-labeled mimecan and DIG-labeled TH antisense probes, confirming mimecan expression (red) primarily within the adrenal cortex, while TH (blue) was found in the medulla; no co-expression was observed (image obtained under visible light at 10× magnification). (F) The field shown is the same as (E) under fluorescent light. (G) Sections of adrenal glands hybridized with fluorescein-labeled mimecan and DIG-labeled ADREAM antisense probes, confirming mimecan expression (red) primarily within the adrenal cortex. Mimecan and ADREAM co-expression was observed in the medulla (image obtained under visible light). (H) The field shown is the same as (G) under fluorescent light. (I) Sections of formalin-fixed, paraffin-embedded adrenal glands immunostained with rabbit anti-human mimecan antibody and anti-rabbit IgG antibody. Positive (brown) staining was found primarily in the adrenal cortex. (J) Negative control of (I) with pre-immune rabbit serum used in lieu of mimecan antibody. (K–L) Higher magnifications of (I, J).