Comparison of Muc5AC immunohistochemistry and mRNA expression in the conjunctiva. (A) Representative immunohistochemistry image showed marked decrease of immunostaining in the EDES+/Treatment− group. Fluorescein isothiocyanate (green) and 4’,6-Diamidino-2-phenylindole (DAPI) (blue) were used for Muc5AC and nuclear staining respectively. (B) The Muc5AC mRNA expression (ΔCt) in the EDES+/Treatment− group was significantly lower than the HMWHA, DQ, and EDES−/Treatment− groups. In addition, the Muc5AC mRNA expression in the HMWHA group was significantly higher than the 0.1% LMWHA, 0.3% LMWHA, and EDES+/Treatment− groups. * and ** represent p < 0.05 and p < 0.001, respectively. LMWHA, low molecular weight hyaluronic acid (HA); DQ, diquafosol sodium; HMWHA, high molecular weight HA; EDES, environmental dry eye stress.