Journal: Cell Reports

Article Title: Silc1 long noncoding RNA is an immediate-early gene promoting efficient memory formation

doi: 10.1016/j.celrep.2023.113168

Figure Lengend Snippet: 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 .

Article Snippet: 8 weeks-old C57BL/6J male mice (Envigo, Israel, n = 3 per group) received bilateral stereotaxic injections of AAV9 Silc1, AAV9 Sox11 or AAV9 GFP control into the hippocampus CA3 regions (titer of 10 12 vg/mL, 0.2 μL Min).

Techniques: Expressing, Control, RNAscope, Immunostaining, Imaging, Injection, Staining, Western Blot

Journal: Cell Reports

Article Title: Silc1 long noncoding RNA is an immediate-early gene promoting efficient memory formation

doi: 10.1016/j.celrep.2023.113168

Figure Lengend Snippet: OE of Sox11 and Silc1 in the hippocampus (A) AAV9-Silc1, AAV9-Sox11, or AAV9-GFP was injected into the CA3 region for Silc1 or Sox11 OE. After 2–3 weeks, the hippocampus was extracted for coronal sections. RNAscope analysis of Silc1 and Sox11 expression using Silc1 (green) and Sox11 CDS (red) probes and DAPI. Imaging was done using 20× (scale bar, 200 μm) and 100× oil immersion objectives (scale bar, 20 μm). (B) RNA-seq quantification of Silc1 and Sox11 ; 3 biological repeats. Mean ± SEM is shown. The p values were calculated using unpaired two-sample t test; ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.001. (C) Immunostaining with anti-SOX11 (red) and DAPI (blue) in hippocampi of Silc1 and Sox11 OE mice. Imaging was performed using a 20× objective (scale bar, 200 μm). See also Figure S4 .

Article Snippet: 8 weeks-old C57BL/6J male mice (Envigo, Israel, n = 3 per group) received bilateral stereotaxic injections of AAV9 Silc1, AAV9 Sox11 or AAV9 GFP control into the hippocampus CA3 regions (titer of 10 12 vg/mL, 0.2 μL Min).

Techniques: Injection, RNAscope, Expressing, Imaging, RNA Sequencing, Immunostaining

Journal: Cell Reports

Article Title: Silc1 long noncoding RNA is an immediate-early gene promoting efficient memory formation

doi: 10.1016/j.celrep.2023.113168

Figure Lengend Snippet:

Article Snippet: 8 weeks-old C57BL/6J male mice (Envigo, Israel, n = 3 per group) received bilateral stereotaxic injections of AAV9 Silc1, AAV9 Sox11 or AAV9 GFP control into the hippocampus CA3 regions (titer of 10 12 vg/mL, 0.2 μL Min).

Techniques: Virus, Recombinant, RNAscope, Multiplex Assay, DNA Library Preparation, Expressing, Plasmid Preparation, Software