Extended Data Fig. 4: Hippocampal downregulation of core-PC proteins does not affect exploratory- and anxiety-like behaviors.

A) As control, preference index for the (left versus right) object location during the training phase of the NOR was measured in 3-month-old mice, 3 weeks after hippocampal stereotactic injections with either AAV-U6-shRNA-Ift20, AAV-U6-shRNA-Kif3a, or their respective AAV-U6-shRNA-Scramble. The NOR was performed in two independent experiments for each group and its respective control. B-C) Open field test (OFT) (B) and Light-dark test (LDT) (C) performed in 3-month-old mice, 3 weeks after hippocampal stereotactic injections with either AAV-U6-shRNA-Ift20, AAV-U6-shRNA-Kif3a, or their respective AAV-U6-shRNA-Scramble. For the OFT, time spent in the center of the arena (%), total ambulation (meter), and speed (cm/s) were measured. For the LDT, number of entries in the light compartment, latency to reach the light box (seconds) and distance spent in the light box (%) were measured. These analyses were performed on 1 cohort of mice for each group. D) Representative western blot and quantification of IFT88 in mouse hippocampi, 3 weeks after stereotactic injections with either eSYN-shRNA-Ift88-mir or eSYN-shRNA-Scramble-mir. β-actin was used as a loading control for each sample. This analysis was performed on 2 independent cohorts of mice for each group. E) Contextual Fear Conditioning test (CFC) performed in 3-month-old mice, 3 weeks after stereotactic injections with either eSYN-shRNA-Ift88-mir or eSYN-shRNA-Scramble-mir. Percent freezing was measured for the training and testing phases. The CFC was performed in 2 independent cohorts of mice. F-G) Preference index for the (left versus right) object location during the training phase of the NOR (F), and number of entries in the light compartment and latency to reach the light box (seconds) during the Light-dark test (LDT) (G) performed in 3 month-old mice, 3 weeks after hippocampal stereotactic injections with either eSYN-shRNA-Ift20-mir or eSYN-shRNA-Scramble-mir. Data are expressed as mean ± SEM.The p values were determined by a two-tailed Student’s t-test compared to U6-shRNA-Scrb in B and C or to eSYN-shRNA-Scrb in D, E and G.

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