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. 2019 Sep 9;9:222. doi: 10.1038/s41398-019-0558-7

Fig. 4. Neonatal Btbd3 knockdown in whole hippocampus recapitulates the Btbd3 knockout phenotype.

Fig. 4

Cre virus successfully knocked down RNA (a, top left) and protein (a, top right; bottom) in hippocampus in a qPCR assay (n = 7 AAV-tdTomato, 2 AAV-Cre virus) and a western blot (n = 3/virus) relative to CTRL, respectively. The western blot image (a, bottom) shows samples from CTRL virus-infused mice in lanes 1, 3, and 5, and samples from Btbd3 KD mice in lanes 2, 4, and 6. The dotted line indicates that the image was interrupted between these lanes in order to remove space between the lanes, but the left and right-hand sides are from the same image of the blot. b Representative example of viral reporter tdTomato expression in hippocampus. c Hippocampal Btbd3 KD did not affect number of wheel revolutions in the voluntary home cage wheel-running assessment (n = 19 female/25 male AAV-tdTomato and 15 female/12 male AAV-Cre infused mice). d Hippocampal Btbd3 KD increased distance traveled in the OF throughout testing (n = 19 female/26 male AAV-tdTomato and 16 female/13 male AAV-Cre infused mice). e Hippocampal Btbd3 KD reduced instances of vertical rearing only within the first 5-min bin. f Hippocampal Btbd3 KD reduced time spent rearing in the first half of testing. g Hippocampal Btbd3 KD reduced spatial d throughout testing (example traces in h, i). Hippocampal Btbd3 KD mice buried fewer marbles in the marble-burying paradigm (n = 19 female/26 male AAV-tdTomato and 16 female/13 male AAV-Cre infused mice) j and left more nestlet unused in the overnight nest-building test k than CTRL mice (n = 19 female/26 male AAV-tdTomato and 16 female/13 male AAV-Cre infused mice). Results are expressed as mean values ± SEM. *p < 0.05 vs. CTRL group in ANOVAs resolved with post hoc tests. KD knockdown, CTRL control virus: AAV-tdTomato, OF open field, AAV adeno-associated virus, qPCR quantitative polymerase chain reaction, Cre AAV-Cre virus