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. 2024 Mar 19;29(8):2496–2509. doi: 10.1038/s41380-024-02515-7

Fig. 6. Restoration of ETV5 in the mPFC rescues impaired sociability in Rfwd2+/− male mice.

Fig. 6

A Experimental design involving injection of the AAV9-CMV-ETV5-eGFP encoding ETV5-eGFP (AAV-ETV5) or empty vector AAV9-CMV-eGFP encoding EGFP only [AAV-CTRL (control), empty vector control without ETV5] into the bilateral mPFC of Rfwd2+/− mice (n = 6, 60 days old); behavioral tests were conducted 3 weeks post-injection. 3CT: 3-chamber test. OFT: open field test. B Representative confocal image showing the expression of AAV-CMV-ETV5-eGFP in the mPFC. C Representative traces of an ETV5-expressing or vector control Rfwd2+/− mouse in the OFT. D–F The ETV5-expressing mice showed no alterations in locomotion (D), time spent in the center (E), or rearing (F) in the OFT compared to controls. G–O Expression of ETV5 rescued impaired social behavior in the 3-chamber test in Rfwd2+/− male mice. G Habituation to the three-chamber apparatus. ETV5-expressing and vector control mice showed similar amounts of time spent in the empty cages (H) and preference index for two empty cages (I). J Schematic of the sociability test. ETV5-expressing and vector control Rfwd2+/− mice showed similar interaction times with mouse 1 (K) and preference index (L) for an unfamiliar mouse 1 (M1), but ETV5-expressing mice showed increased interaction time with a novel mouse (N) and increased preference index (O) compared to control mice. M Representative traces from an ETV5-expressing or vector control mouse in the ‘Familiar Mouse1–novel mouse’ test. P–W Expression of ETV5 did not rescue the spatial memory deficit in Rfwd2+/− mice. P Experimental diagram of the Barnes maze, and representative traces of the probe trials on days 6 and 9. Q Escape latency during training trials on days 2–5. Latency to locate the escape hole (R, S), error entries to locate the target (T, U), and time spent in the target quadrant (V, W) 24 h and 4 days after training on days 6 and 9 were not different between ETV5-expressing and control mice, respectively. H, K, N two-way ANOVA followed by Bonferroni’s test. Q Two-way repeated-measures ANOVA followed by Sidak’s test, others with two-population Student’s t-test. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01, ***p < 0.001.