Morphological assessment of ethanol-induced apoptosis via TUNEL assay. (A) Cells were treated for 12 h with normal media as control (C), ethanol (EtOH, 100 mM), TQ (25 μM) plus ethanol (TQ+EtOH), Met (10 mM) plus ethanol (Met+EtOH), TQ plus Met plus ethanol (TQ+Met+EtOH), respectively. Effects of ethanol, TQ plus ethanol, Met plus ethanol and TQ plus Met plus ethanol on apoptotic death in prenatal rat cortical neurons was visualized with TUNEL and DAPI stains. Representative photomicrographs of TUNEL staining show apoptotic neurons after ethanol administration followed by TQ and Met. TQ and Met treatment effectively blocked ethanol-induced apoptosis, as evidenced from the lack of TUNEL-positive cells. Panels A-O display TUNEL-stained cells observed by confocal microscopy at higher magnifications with a 40× objective field, Scale bar = 20 (B): The percentage of TUNEL positive cells in each case were counted and the cumulative data from three independent experiments is shown here as mean ± SEM (n = 3). #P < 0.05 significantly different from control; *P < 0.05 different from ethanol.