Consistent with previous studies showing cross talk between the inflammasome pathway and lysosomal-mitochondrial machinery [8], we investigated the mitochondrial and lysosomal patterning in iPS cells and NPCs with or without ethanol exposure, as well as in neurons derived from NPCs with or without ethanol pre-exposure. In particular, we investigated the distribution of mitochondria and lysosomes using a Mitotracker Assay™ (Fig. 4) and IHC for the lysosomal marker Lamp1 (Fig. 5), respectively. Both mitochondria (Fig. 4) and lysosomes (Fig. 5) appear to be reduced in iPS cells or NPCs exposed to ethanol. In contrast to the more dispersed mitochondrial distribution in untreated iPS cells, NPCs, or neurons, the mitochondria cluster more prominently in the perinuclear area in treated cells, with a particularly robust effect in the 7d condition (Fig. 4a, b and 5a, b). Similar effects were observed in neurons derived from NPCs that were pre-exposed to ethanol (Fig. 4c and 5c).
