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. 2015 May 13;194(12):5990–5998. doi: 10.4049/jimmunol.1401489

FIGURE 1.

FIGURE 1.

Vinpocetine inhibits S. pneumoniae–induced mucin MUC5AC expression in vitro. HMEEC were pretreated with vinpocetine [5, 10, and 20 μm in (A), 10 μM in (B)–(G)] for 1 h, followed by stimulation of S. pneumoniae [5 h in (A)–(D), or 12 h in (E)–(G)]. Relative quantity of MUC5AC mRNA expression was measured with real-time qPCR analysis in (A) HMEEC, (B) HMEEC cultured under air-liquid interface conditions, or (C) HMEEC stimulated with S. pneumoniae strains 6B, 19F, 23F, or D39. (D) HMEEC transfected with MUC5AC-luciferase reporter gene were pretreated with vinpocetine, followed by S. pneumoniae stimulation, and MUC5AC transcriptional activity was measured with luciferase assay. (E) Expression level of MUC5AC protein was measured in the cell culture supernatant with direct ELISA against MUC5AC. (F) HMEEC were stained with Ab against MUC5AC, probed with FITC-conjugate, and (G) MUC5AC expression in IF-stained cells was analyzed by a relative quantification of immunofluorescence (original magnification ×200; scale bars, 20 μm). Data in (A)–(E) and (G) are mean ± SD (A–E, n = 3; G, n = 5). Statistical analysis was performed using Student t test. Pictures from IF-stained cells of one representative experiment are shown in (F) (n = 5). Data are representative of three or more independent experiments. *p < 0.05. CON, control; Sp, S. pneumoniae; Vinp, vinpocetine.