The inflammasome pathway is known as an alternative to apoptosis, leading progressively to cell degeneration and death. Ethanol has been shown to activate this pathway in different cell systems [17, 24, 25], including mouse astrocytes [5], thus demonstrating the ability of neural cells to participate in the development of an inflammatory cascade. Given our data in iPS cells suggesting ethanol-induced cellular inflammation (Fig. 1), we sought to determine if ethanol activates the inflammasome pathway in NPCs. IHC analysis in NPCs revealed a robust increase of the number of Casp1+ cells after ethanol exposure, with no significant difference between 24hr vs. 7d exposure (Fig. 2f and g). Western Blot analysis revealed an increase in the expression of both the prodomain (p45) and the activated domain (p10) of Casp1 (Fig. 2h). This effect mirrored a similar increase in NLRP3 expression (Fig. 2f and i), thus confirming the involvement of the inflammasome pathway. Since the inflammasome-activated pathway is known to interplay with autophagy [6, 8, 9], we also evaluated the distribution of LC3B (a marker of the autophagosome membrane [26]) puncta in NPCs.
