Real-time quantitative RT-PCR for ZO-1 mRNA in the ALF groups and other control groups. ZO-1 mRNA level was decreased at 9 h in mice with ALF. Total RNA derived from each tissue was reverse transcribed and subjected to real -time quantitative RT-PCR to evaluate ZO-1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA levels. Results are expressed as the number of ZO-1 amplicons per 104 GAPDH amplicons. Bars represent the mean value for each group. The only significant difference in ZO-1 mRNA level between the ALF and control groups (NS: 100.00 ± 0.00%) was at 9 h. At this time, there was decrease in the level of full-length ZO-1 mRNA in the ALF mice. At all other times, there was no difference between the ALF group and the other groups, which is consistent with our immunohistochemistry results.