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Inducible nitric oxide synthase (NOS‐2) is abundantly present in the optic nerve heads of glaucoma patients. To determine the regulation of NOS‐2 expression in the glaucomatous optic nerve head, the specific cells that express NOS‐2 in the optic nerve heads of patients with primary open‐angle glaucoma were studied by immunohistochemical double‐labeling of NOS‐2 and one of the characteristic cell markers for different cell types. Most of the labeling for NOS‐2 was identified in reactive astrocytes that were clustered in the areas of nerve damage in the prelaminar and lamina cribrosa regions of the glaucomatous optic nerve heads. In vitro, the expression of GFAP and NOS‐2 by reactive astrocytes of human optic nerve heads was demonstrated by immunocytochemistry and Western blot. In primary cultures of human lamina cribrosa astrocytes, stimulation by interferon‐γ and interleukin‐1β upregulated GFAP …