Figure 2.

Expression of Silc1 and Sox11 under control and novel environment (NE) conditions in WT and Silc1^−/− mice

(A) RNAscope FISH assay on hippocampal sections from mice with the indicated genotype. Tissues were counterstained with a Silc1 probe (red) and DAPI (blue) and imaged using 20× (scale bar, 200 μm) and 100× oil immersion objectives (scale bar, 20 μm). NE exposure was performed using the Barnes maze setting, and the hippocampus was extracted for coronal sections after 1 h of stimulus. 12 images of non-overlapping fields per biological repeat were quantified; 3 biological repeats. Mean ± SEM is shown. The p value was calculated using unpaired two-sample t test; ^∗∗p < 0.005.

(B) As in (A) with tissues hybridized with Sox11 CDS (red) and 3′ UTR (green) probes and counterstained with DAPI (blue).

(C) As in (A) for the number of green and red dots. The p value was calculated using unpaired two-sample t test; ^∗p < 0.05.

(D) Immunostaining with anti-SOX11 (red) and DAPI (blue) in hippocampi of WT and Silc1^−/− mice under HC and NE conditions. Imaging was performed using a 20× objective (scale bar, 200 μm).

(E) As in (A) for Sox11^fl/fl mice injected stereotaxically in the CA3 region with Cre-GFP- or GFP-expressing AAV9 viruses. Imaging was done using a 20× objective (scale bar, 200 μm). SOX11 levels were significantly reduced after Cre injection, which indicates the specificity of the SOX11 antibody used for staining.

(F) Quantification of 3 biological repeats of hippocampus staining. Mean ± SEM, ^∗p < 0.05, ^∗∗p < 0.005, unpaired two-sample t test.

(G) Western blot with SOX11 and β-tubulin antibodies from mice with the indicated genotype, using whole-hippocampus protein extract. Hippocampus protein extract from Sox11^fl/fl mice that were stereotaxically injected in the CA3 region using AAV9 Cre-GFP or AAV9 GFP were used as a specificity control for the antibodies.

(H) Western blot quantification. SOX11 expression levels were normalized to β-tubulin levels. n = 3. Mean ± SEM is shown; ^∗p < 0.05, unpaired two-sample t test.

(I) As in (A) for hippocampus sections from Silc1^fl/fl mice that were stereotaxically injected in the CA3 region using AAV9 Cre-GFP or AAV9 GFP. Two weeks after injection, the mice were exposed to an NE, and the hippocampus was extracted for coronal sections after 1 h of NE exposure. Imaging was performed using a 20× objective (scale bar, 200 μm).

See also Figure S2.