Fig. 3

Early ethanol exposure leads to a decrease of NPC-derived neurons. NPCs were pretreated with ethanol for 24hr or 7d and differentiated to neurons for 26 days. a Phase contrast pictures of treated and untreated NPCs after 7 days of differentiation toward the neuronal lineage, and immunofluorescence analysis of neurons differentiated from NPCs showing the synaptic marker synapsin, and glutamatergic marker vGlut. Scale bars: 200 μm (top), 10 μm (middle, bottom). b Graph showing the relative percentage of MAP2⁺ cells over the total number of DAPI⁺ nuclei. c Number of synapses quantified by synapsin⁺ puncta per 100 μm dendrite length under control or ethanol pre-exposure conditions. d Electrophysiological analysis. e Immunofluorescence analysis showing the expression of Casp1, MAP2, and NLRP3. Scale bar: 50 μm. The differences among all the values were not statistically significant unless indicated (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). Student’s t-test was utilized for all experiments