Fig. 1

Ethanol activates the inflammasome pathway in iPS cells. iPS cells were cultured in 70 mM ethanol for 24hr or 7d. a From the top: Immunofluorescence analysis by confocal microscopy shows the expression of pluripotency markers Oct3-4, Tra-1-60, Sox2, and of the proliferation marker Ki-67 in iPS cells after 24hr or 7d ethanol exposure. Scale bar: 50 μm. b Growth curve of iPS cell lines #1 and #2. Inserts: showing the doubling time (days in vitro) (left), and the relative percentage of pyknotic nuclei over total DAPI⁺ nuclei (right). c Confocal microscopy images showing the expression of the apoptotic marker, Cleaved Caspase-3 (Casp3) and of the inflammasome-related markers, Caspase-1 (Casp1) and NLRP3. Scale bars: 50 μm. d Western Blot analysis and graph showing the relative densitometric analysis of expression of the inflammasome-pathway markers Casp1 (p45 and p10) and NLRP3. Quantification of the proteins was normalized to β-actin expression. e Graph showing the relative percentage of Casp3⁺ cells over the total number of DAPI⁺ cell nuclei. f Western Blot analysis of Casp3 expression