Effects of EtOH and acamprosate on iPSC-derived astrocytes. (a) Down-regulation of TSPAN5 mRNA expression (F4,30 = 24.25, p = 0.0007) and (b) 5-HT concentrations (F4,35 = 4.46, p = 0.046) in iPSC-derived astrocyte culture medium (n=5) in the presence of EtOH (from 5 mM to 50 mM) for 24 hours. (c) Down-regulation of TSPAN5 mRNA expression (F4,35 = 4.54, p = 0.037) and (d) 5-HT concentrations (F4,35 = 4.65, p = 0.031) in iPSC-derived astrocyte culture medium (n=5) in the presence of acamprosate (0.25 μM to 10 μM) for 24 hours. (e) Knockdown of TSPAN5 (f) decreased the 5-HT concentrations in iPSC-derived astrocyte culture medium. Five independent human-derived-iPSC-derived astrocyte cell lines were used to perform these experiments. Cells were seeded in T75 culture flasks at a density of 2.5 × 106 cells. *A p value ≤0.05 was considered statistically significant. Three independent experiments were performed. All values are mean±SEM.