Fig. 3: Genetic deletion of AMPK in the mPFC elicited anxiety-like behaviors and abolished the anxiolytic effect of metformin.

a rAAV2/8 virus expressing Cre recombinase (AMPK KO^mPFC) or EGFP control (AMPK Flox) was microinjected into the mPFC of adult AMPK Flox mice. Representative viral expression (green). Scale bar, 100 μm. b Western blotting of the mPFC and (c) related quantification; n = 3 mice. d In the LDT, the residence time in the light box (left) and the latency for mice to enter the light box from the dark box (right). e In EPM, the entry times in the open arms (left), the time in closed arms (middle), and the total entries in the open and closed arms (right). In (c, d, e), unpaired Student’s t test, n = 9 or 11 mice. f–h AMPK KO^mPFC and AMPK Flox mice were subjected to RSD stress and metformin treatment (i.g., 250 mg/kg) as mentioned. Veh, vehicle; Met, metformin. f In the EPM, the time spent (left) and the distance traveled (right) in the open arms. In the OFT, (g) representative traces of mice and (h) the time (left) and entries (right) in the center area. In (f, h), two-way ANOVA followed by Bonferroni’s test, n = 10–13 mice. *P < 0.05, **P < 0.01.