Figure 1.

Experimental schematic and acute vs. prolonged Tat expression. C57BL/6 wild-type (WT) and iTat mice of both sexes were administered doxycycline (DOX) via intraperitoneal (i.p.) injections (100 mg/kg/day) or DOX-containing food ad libitum (about 5–6 mg DOX/day). (A) A total of seven i.p. injections were given to 5–7 month old mice over 3 weeks prior to and during behavior testing. In parallel, mice were fed DOX food for about 7 months (6 months prior to and 1 month during the behavior testing) starting at 4 to 5 weeks of age. Therefore, these groups were age-matched at the initiation of behavior testing, which was carried out over 4 weeks. After behavior testing, mice were euthanized, and brain tissues were harvested for gene expression and/or protein analyses. (B) Tat mRNA expression was measured in iTat mice using one-step real-time PCR. GAPDH was used as an internal housekeeping control. The iTat mice received acute Tat induction by DOX i.p. injections (n = 11, open squares) and prolonged Tat induction via DOX food (n = 13, solid squares). Each bar represents the mean ± SEM. ^#p < 0.050 by unpaired T-test. (C) Tat protein expression was measured in WT and iTat mice using simple western (WES, protein simple). GAPDH was used as an internal housekeeping control. The WT and iTat mice received acute Tat induction by DOX i.p. injections (n = 10, open squares) and prolonged Tat induction via DOX food (n = 7, solid squares). Each bar represents the mean ± SEM. ^#p < 0.050 by unpaired T-test.