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After the test, the number of transitions across area boundaries and time spent in the four central areas were recorded from video observation. The EPM was made of gray-painted wood and consisted of a 60 �� 60 mm center platform and four [https://en.wikipedia.org/wiki/Glafenine Glafenine] arms (60 �� 300 mm) extending from the platform in a cross formation, with two opposing arms enclosed by side walls (300 �� 150 mm) and two open arms. The entire maze was elevated 30 cm above the floor and illuminated from above by a dim red light. At the beginning of the test, a mouse was placed in the center platform facing one of the open arms and behavior was recorded for 5 min by a camera attached approximately 100 cm above the apparatus. The number of entries into and time spent in the open arms and closed arms were recorded from video observation. We analyzed the time in the open arms to total arm time (%) and the number of entries into these arms to total arm entries (%) as indices of anxiety-like behavior and the number of entries into the closed arms as an index of activity. The PAL test apparatus was a rectangular Plexiglas chamber consisting of two compartments, one white and the other black, separated by a common wall (Takei Scientific Instruments Co., Ltd., Niigata, Japan). On day 1, a conditioning trial was conducted. Subjects were placed in the compartment with white walls, and the door into the compartment with black walls was opened 3 min later. After the mice entered the dark compartment, a 0.7 ��A shock was applied to the floor of the dark chamber for 3 s. The subjects were returned to their home cages 1 min after shock delivery. The next day, [http://www.selleckchem.com/pharmacological_MAPK.html p38 MAPK signaling] subjects were placed individually in the light compartment for the test trials. Six seconds after introduction, the door was opened and their behavior was monitored for 180 s or until the subject crossed into the dark compartment. The latency (s) to cross was recorded for each trial. Statistical analyses All data are presented as mean �� standard error (SE). Two-sample t-tests were used to compare means between DES and OIL-control groups. When the data did not fit a normal distribution, the Mann�CWhitney U-test was used as an alternative. Two-Way ANOVAs were used when two factors were analyzed such as maternal and offspring exposure to DES. When [http://www.selleckchem.com/products/E7080.html Lenvatinib cost] necessary a test of a simple main effect was conducted to estimate the group difference at each level. Ethics All experimental procedures were in strict accordance with the guidelines of the Care and Use of Laboratory Animals in Kagoshima University and approved by the Ethics Committee for Animal Experimentation at Kagoshima University. Results Effects of DES on maternal behavior DES exposure during gestation reduced subsequent maternal behavior by dams in the undisturbed condition. Mothers fed with 0.1 ��g DES/day (DES group) showed decreased levels of arched-back posture [U(10/9) = 13, p