EMBO/soda-nel huggingface dataset:
Oligonucleotide-induced alternative splicing of serotonin 2C receptor reduces food intake (2016)
Figure 4

Figure 4: Oligo#5 influences the ratio of the two serotonin 2C receptor isoforms and changes the localization of the full-length receptor

A. Structure of the reporter gene introduced into cells. The start codon in exon III is shown (circle) and the splicing patterns emerging from using the distal and proximal splice site (DS, PS). Skipping of exon Vb causes a frameshift, which terminates the protein in the first stop codon (square). Exon Vb inclusion keeps a longer open reading frame, that terminates after the GFP sequence. The encoded proteins are schematically shown below.

B. Effect of oligo#5 on pre-mRNA splicing of the reporter gene, stably transfected into HeLa cells.

C. Effect of oligo#5 on the truncated 5HT2C protein. Stable cell lines expressing the construct shown in panel (A) were treated with oligo#3 and oligo#5. After 48 hrs cells were separated in membrane containing fractions and soluble cytosolic supernatant. The fractions were analyzed by Western Blot using the anti-RNA1 antiserum (Figure EV1). The protein samples were on the same membrane, but the supernatant fractions were exposed about ten times longer.

D. Effect of oligo#5 on the full-length 5HT2C protein. The protein was prepared as in panel (C) and analyzed using a polyclonal anti-GFP antiserum.

E. Quantification of panels C and D from three independent experiments. The ratio of protein signal after oligo#3 and oligo#5 treatment is shown for RNA1 (p=0.002) and RNA2 (p=0.0003), n=3, respectively.

F, G. Effect of oligo#5 on localization of the EGFP-tagged serotonin 2C receptor. Confocal images of three representative pictures are shown.

H. Quantitification of GFP signal found on the membrane after oligo#5 addition. The signal around the peripheral region was determined by visual inspection and calculated as a fraction of total signal using NIH image. 200 cells from three individual experiments were analyzed, p<0.008, n=200.
Oligonucleotide-induced alternative splicing of serotonin 2C receptor reduces food intake (2016)
Figure 5

Figure 5: Oligo#5 enters neurons after ICV injection, changes 5HT2C splicing and food intake

A. Accumulation of Cy3-labeled oligo#5 in the area around the 3rd ventricle one hour post injection, shown is the ventral part of a coronal section -1.34 mm from Bregma. The injection coordinates were AP: -1.82 mm, DV: -4.5 mm, ML: 0.

B. Uptake of oligo#5 in the arcuate nucleus region (arc), enlarged from panel A.

C. Oligo#5 enters neurons. Enlargement of hypothalamus region taking up oligo#5 (red), stained for the neuronal marker NeuN (green). Most of the oligo#5 signal is in neurons, but some is also in NeuN negative cells (arrow), likely glia.

D. Oligo#5 enters some glia cells. Enlargement of a hypothalamic region adjacent to the 3rd ventricle one hour after oligo#5 injection. The oligo (red) can be found in some GFAP-positive glia cells (arrow), but numerous GFAP-positive cells (green) show no oligo#5 uptake.

E. Effect of oligo#5 on RNA1/RNA2 ratios in the injected area. The area marked by Cy3 staining in the dorsal hypothalamus, consisting mainly of the arcuate nucleus was microdissected, RNA isolated and analyzed by qPCR. NS: non-significant changes (3 hrs: p=0.01; 6 hrs: p=0.000001; 9 hrs: p=0.02; 12 hrs: p=0.06; n=3)

F. Oligo#5 does not change the RNA2-editing pattern. The hypothalamic area showing oligo#5 staining was microdissected 6 hrs post injection and RNA2 was amplified and the PCR product subjected to deep sequencing. The percent of non-edited reads in total reads is shown (at least 20,000 reads per lane).

G. Effect of oligo#5 on POMC expression. The RNA from panel D was analyzed using qPCR with primers against POMC, normalized to GAPDH (3 hrs: p=0.0009; 6 hrs: p=0.02; 9 hrs: p=0.1; 12 hrs: p=0.3, n=3).

H. Effect of oligo#5 on food uptake in wild-type mice. C57BL/6 wild-type mice received a guide canulae and did not have access to food for 16 hours. After injection of 2 µg oligonucleotides, they could freely access food. The food consumption was measured by weighing the food used. A total of 52 animals were used, p=0.001 for the difference between oligo#5 and control oligonucleotides. Each of the six individual experiments showed statistically significant differences between oligo#5 and control (p<0.01 – 0.001). Control oligo: an oligonucleotide against human SMN2.

I. Effect of oligo#5 on food uptake in ob/ob mice. Ob/ob mice received oligo#5 through ICV injection, similar to panel F. However, the mice could freely access food at all times (p=0.04, n=3).
Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding (2021)
Figure 1

Fig 1 Molecular architecture of yeast Smc5/6 complexes determined by cross-linking mass-spectrometry (XL-MS) and cryo-electron microscopy (cryo-EM).

A. Reconstitution of Smc5/6 and Nse5/6 complexes. Left panel, schematic depiction of the composition and organization of the yeast Smc5/6 'core' hexamer and the Nse5/6 dimer. Middle panel, elution profiles for analytical gel filtration (Superose 6 Increase 3.2/300) of the Smc5/6 core hexamer, the Nse5/6 dimer and a holo-complex obtained by mixing of dimer and hexamer. Measured by absorption at 280 nm. Right panel, peak fractions were analysed by SDS-Page and Coomassie Brilliant Blue staining.

B. Circular representation of lysine-lysine inter-subunit cross-links identified by mass spectrometry (XL-MS) in buffer containing 250 mM NaCl. For simplicity, cross-links between proteins (or domains) are grouped, and the thickness of the lines indicate the total number of crosslinks of this particular type. Smc5 and Smc6 proteins are divided into N- and C-terminal head domains (HD) and coiled-coil arms (CC) as well as the central hinge domain. For a full representation of individual inter-subunit as well as intra-subunit cross-links, see Dataset EV1.

C. Cross-links obtained within and between Smc5 and Smc6 subunits of the Smc5/6 octamer. A cross-link that did not match to the elongated rod-shaped particle is displayed by a dashed line in red colours. The cartoon on the right shows the dimer of folded Smc5 and Smc6 proteins, with examples of intra- and interlinks within the coiled-coil arms indicated with yellow and red lines, respectively.

D. Selected 2D class averages obtained by cryo-electron microscopy of the yeast Smc5/6 hexamer. Representative classes with dimers of Smc5/6 hexamers are displayed (left images). Emerging details are indicated schematically (right panel).
Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding (2021)
Figure 2

Fig 2 A Nse5/6 co-crystal structure.

A. Domain organization of Nse5 and Nse6 proteins. Putative domain boundaries identified by secondary structure prediction and limited proteolysis are denoted by arrowheads, in black colours when producing stable fragments, otherwise in grey colours.

B. Limited proteolysis of a purified preparation of the Nse5/Nse6 complex by trypsin. Samples taken at different time points were analyzed by SDS-Page and Coomassie staining. Selected stable fragments identified by mass spectrometry are indicated.

C. Front and back view of the Nse5/Nse6 co-crystal structure in cartoon representation. Structural elements of Nse5 and Nse6 are displayed in orange and purple colours, respectively. α-helices are labelled.

D. Conservation of residues at the Nse5/6 binding interface. Interaction surface of Nse6 (left panel) and Nse5 (right panel). For orientation, the secondary structure at the interface is displayed (middle panel). Colour code for residue conservation is given at the bottom of the panel.

E. A conserved surface area (bottom panel) on top of Nse5/6. Colour coding for conserved residues as in (D). An extended loop in this region contains a lysine residue (K148) that was found to crosslink to Smc5 and Smc6 ATPase heads (see Fig. 3).
Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding (2021)
Figure 3

Fig 3 Association of the yeast Smc5/6 hexamer with the Nse5/6 dimer.

A. Pulldown ('pd') assays using immobilized Smc5/Smc6-Twin-Strep ('TS') hexamers and soluble input material ('in') of Nse5/6 (left), of Nse5/Nse6(177-C) (middle) and of Nse6(1-179)-CPD-His (right). Control pulldowns ('-') were performed by omitting the pre-binding of Smc5/Smc6-Twin-Strep to the beads. Fractions were analysed by SDS-Page and Coomassie staining.

B. Salt stability of Smc5/Smc6-Twin-Strep interactions with Nse5/6 and Nse6(1-179). Immobilized Smc5/Smc6-Twin-Strep was mixed with Nse5/6 or Nse6(1-179)-CPD-His ('in'). Beads were washed with buffers containing the indicated salt concentrations. Bead fractions were analysed by SDS-Page and Coomassie staining. Pulldown efficiencies were estimated from the intensity of Coomassie gel bands.

C. XL-MS cross-links detected between Nse5 and Nse6 proteins and subunits of the Smc5/6 hexamer in schematic representation (middle panel). The positions of lysine residues on the Smc6 head and the Smc5 head, left and right panels, respectively, with cross-links to Nse5 are denoted on Phyre2-generated homology models.

D. Analysis of spore viability by yeast tetrad dissection. Diploid strains heterozygous for alleles of Nse6 (wt, 86-C, 177-C and 179-C) were sporulated. Isolated spores were grown on YPD plates. Viable clones were tested for the marker cassette conferring resistance to G418 (marked by circles in green colours). Dead spores were marked by circles in red colours.
Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding (2021)
Figure 4

Fig 4 ATP hydrolysis by purified Smc5/6 hexamer and octamer.

A. ATP hydrolysis rates (given per Smc5/6 complex) were measured by an enzyme-coupled assay in the absence and presence of plasmidDNA (conc. 1.25 nM) for the Smc5/6 hexamer (conc. 150 nM) and the octamer (conc. 150 nM) with increasing concentrations of ATP. The curves were fitted to Michaelis Menten equation and Km and kcat values were determined. Please note that the hexamer without plasmidDNA shows cooperative behaviour, thus the Michalis-Menten kinetics is not formally applicable (marked by asterisk). Assays were performed in biological triplicates, mean values are shown with error bars indicating standard deviations. For bar graphs individual data points are also displayed.

B. Same as in (A) using 40bpDNA (annealed 40 bp oligo DNA) (conc. 1 µM) instead of plasmidDNA. Note that the data points and curves for samples without DNA (hexamer -DNA; octamer -DNA) are identical to (A).

C. Fluorescence anisotropy measurements using 40 bp dsDNA or 40mer ssDNA substrates. Representative binding curves (top graph) and the resulting Kd values (bottom graph) indicate that both hexameric and octameric Smc5/6 complexes interact with both substrates, whereas the Nse5/6 complex alone does not. The bar graph shows mean values with standard deviations from technical triplicates. Individual data points are also displayed.

D. Pulldown experiments with Smc5/6 complexes and circular plasmid (2.8 kbp). DNA is retained after high salt washes (1 M NaCl) only in the presence of both Nse5/6 and ATP. The graph on the right shows a quantification (mean values and standard deviations) of the amount of co-purified DNA from technical triplicates. Individual data points are also displayed.

E. Salt-stable DNA association requires the DNA substrate to be circular ('circ'). Same experimental setup as in (D), but the plasmid substrate was also linearized by restriction digest ('lin').

F. Salt-stable DNA association requires ATP binding and Smc5/6 head-engagement, but not ATP hydrolysis. Same experimental setup as in (D) using mutant versions of the Smc5/6 hexamer as well as the non-hydrolyzable analogue of ATP, ATPγS, as indicated.
Nse5/6 inhibits the Smc5/6 ATPase and modulates DNA substrate binding (2021)
Figure 5

Fig 5 Detection of Smc5/6 conformations by cysteine cross-linking.

A. Cross-linking of a purified preparation of wild-type Smc5/6 hexamer in the absence and presence of Nse5/6. ATP and plasmidDNA were added as substrates as denoted. Products were analysed by SDS-Page and Coomassie staining. A cross-linked species derived from natural cysteines is indicated by an arrowhead ('XL'). Note that a fraction of Nse4 is converted into a more slowly migrating species, presumably by intra-molecular cross-linking.

B. Cross-linking of engineered variants of the Smc5/6 hexamer. As in (a) using Smc5 and Smc6 cysteine mutants. Schemes indicate the location of engineered cysteines and their expected ability to cross-link in a rod-like and a ring-like conformation. High-molecular weight species were analysed by SDS-Page and Coomassie staining. Wild-type hexamer ('wt') is included as cross-linking control. Species occurring only in the presence of engineered cysteines are labelled by coloured arrowheads. Cross-linking efficiencies were calculated from the intensity of Coomassie-stained bands by comparing the band of the corresponding cross-linked species to the bands of unmodified Smc5 and Smc6. Numbers below the gel quantify the percentage of cross-linked protein species in the displayed gel. Comparable numbers were obtained in at least one additional independent experiment.

C. DNA effects on Smc5/6 arm co-alignment as judged by CC-Cys cross-linking. As in (B) but also including 40bpDNA. Additon of plasmidDNA but not 40bpDNA reduces CC-Cys cross-linking in the Smc5/6 hexamer.

D. DNA effects on head engagement as judged by E-Cys cross-linking. As in (B) also including 40bpDNA. plasmidDNA binding, but not 40bpDNA binding, overcomes inhibition of E-Cys cross-linking by Nse5/6.

E. Effects of 40bpDNA and plasmidDNA on head engagement (as judged by E-Cys cross-linking) of the Smc5/6 complex with wildtype (wt) or hydrolysis-deficient (EQ) heads, in both the presence or absence of the Nse5/6 complex. Cross-linking efficiencies were calculated from the intensity of Coomassie-stained bands by comparing the band of the corresponding cross-linked species to the bands of unmodified Smc5 and Smc6. Numbers below the gel quantify the percentage of cross-linked protein species in the displayed gel.

F. Effects of plasmidDNA on head engagement of the Smc5/6 complex (as judged by E-Cys cross-linking) in the presence of ATP or the non-hydrolyzable analogue ATPγS. Calculations and quantifications as in (E).
HDAC1 Dysregulation Induces Aberrant Cell Cycle and DNA Damage in Progress of TDP-43 Proteinopathies (2020)
Figure 1

Figure 1. Aberrant cell cycle activity correlates with TDP-43 proteinopathies in FTLD-TDP Tg mice.

Representative immunofluorescence (IF) staining of Ki67 and TDP-43 in the regions of frontal cortices from 6-mon-old WT and FTLD-TDP Tg mice. Nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI; upper panel in blue) or neural marker NeuN (lower panel in green). Scale bar: 50 μm. The circled area is emphasized for showing the distribution of immunoreactivity in cell subregions. Scale bar: 15 μm.

Quantification of cells or neurons with Ki67 immunoreactivity and TDP-43 mislocalization from each view of microscope. n = 9 sections per mouse, N = 5 mice per group, data are presented as mean ± SEM, ***p = 0.0007 by t-test.

Representative data of reverse-transcription PCR (left panel) or Western blot (right panel) for cell cycle-related genes and semi-quantification of the expression levels in the frontal cortices and hippocampus from the 6-mon-old WT and FTLD-TDP Tg mice. N = 5 mice per group, data are presented as mean ± SEM (%), statistical analysis by multiple t-test with FDR correction, Q = 1%. *P < 0.05, **P ≤ 0.01; ***P ≤ 0.001 and ****P ≤ 0.0001, exact p values are shown in Appendix Table S1.
HDAC1 Dysregulation Induces Aberrant Cell Cycle and DNA Damage in Progress of TDP-43 Proteinopathies (2020)
Figure 2

Figure 2. DNA damage correlates with TDP-43 proteinopathies in FTLD-TDP Tg mice.

Representative image of comet assay for DNA fragmentation and the quantification of cells with comet tails in the regions of frontal cortices from 6-mon-old WT and FTLD-TDP Tg mice. Scale bar: 50 μm. n = 9 sections per mouse, N = 5 mice per group, data are presented as mean ± SEM, *p = 0.0114 by t-test.

Representative IF staining of γH2AX and TDP-43 in the regions of frontal cortices from 6-mon-old WT and FTLD-TDP Tg mice. Nuclei were stained with DAPI (upper panel in blue) or NeuN (middle panel in green). Scale bar: 50 μm. The circled area is emphasized for showing the distribution of immunoreactivity in cell subregions. Scale bar: 15 μm. Lower panel: quantification of cells or neurons with γH2AX immunoreactivity and TDP-43 proteinopathies from each view of microscope. n = 9 sections per mouse, N = 5 mice per group, data are presented as mean ± SEM, ***p = 0.0008 by t-test.

Representative IF staining of γH2AX and Ki67 in the regions of frontal cortices from WT and FTLD-TDP Tg mice. Nuclei were stained with DAPI (upper panel) or NeuN (middle panel). Scale bar: 50 μm. Subregions, scale bar: 15 μm. Lower panel: quantification of cells or neurons with γH2AX and Ki67 immunoreactivity from each view of microscope. n = 9 sections per mouse, N = 5 mice per group, data are presented as mean ± SEM, ***p = 0.0004 by t-test.
HDAC1 Dysregulation Induces Aberrant Cell Cycle and DNA Damage in Progress of TDP-43 Proteinopathies (2020)
Figure 3

Figure 3. HDAC1 dysregulation in FTLD-TDP Tg mice.

Top, representative western blot data of HDAC1 and TDP-43 in extracts obtained following RIPA fractional extraction in the frontal cortices and hippocampus from 6-mon-old of FTLD-TDP or WT mice. Bottom, semi-quantification of HDAC1 and TDP-43 expression levels. N = 5 mice per group, data are presented as mean ± SEM (%), **** p < 0.0001 by multiple t-tests.

Top, representative western blot data of HDAC1 and TDP-43 in the cytosolic and nuclear extracts. Bottom, quantification of HDAC1 and TDP-43 levels. N = 5 mice per group, data are presented as mean ± SEM (%), *** p = 0.0005 (TDP-43) or 0.0004 (HDAC1); **** p < 0.0001 by multiple t-tests.

IF staining of HDAC1 and SMI-32 in the regions of frontal cortices from WT and FTLD-TDP Tg mice. Scale bar: 150 μm. Blocked area in left bottom picture is showed in a magnified view in the right bottom picture. Scale bar: 50 μm.

Nuclear HDAC1 activity assay from frontal cortices and hippocampus in 6-mon-old of FTLD-TDP and WT mice. TSA: nuclear extracts that were treated with Trichostatin A (TSA, an HDAC inhibitor) as a negative control for HDAC1-transferred fluorescent activity during the HDAC1 activity assay. N = 5 mice per group, data are presented as mean ± SEM (%), *p = 0.0415 by t-test.

Left, representative western blot data of nuclear acetylated-histone H3 (Lys 9/14) and total-histone H3 in 6-mon-old of WT and FTLD-TDP Tg mice. Right, quantification of nuclear acetylated-histone H3/total-histone H3 ratio. N = 5 mice per group, data are presented as mean ± SEM (%), * p = 0.0147 by t-test.
HDAC1 Dysregulation Induces Aberrant Cell Cycle and DNA Damage in Progress of TDP-43 Proteinopathies (2020)
Figure 4

Figure 4. HDAC1 mislocalization correlates with pathogenesis of TDP-43 proteinopathies in FTLD-TDP Tg mice.

Left graph: IF staining of TDP-43 and HDAC1 during progression of TDP-43 proteinopathies in the frontal cortices from the FTLD-TDP Tg and WT mice. Nuclei were stained with DAPI (in blue). Scale bar: 50 μm. The circled area is emphasized for showing the distribution of immunoreactivity in cell subregions. Scale bar: 15 μm. Right histogram: Quantification of co-localized TDP-43 and HDAC1 immunoreactivity in the cytosol or nucleus in the WT or 1-, 6-, and 12-month-old FTLD-TDP Tg mice. n = 9 sections per mouse, N = 5 mice per group, data are presented as mean ± SEM. *Nucleus: Tg 1m vs. Tg 6m or 12m; #Cytosol: Tg 1m vs. Tg 6m or 12m; @ Tg 6m vs Tg 12m. ****/####/@@@@ p < 0.0001 by multiple comparison.

Left graph: IF staining of γH2AX and HDAC1 in the frontal cortices from the 12-month-old FTLD-TDP Tg and WT mice. Nuclei were stained with DAPI (in blue). Arrow head: γH2AX+ nucleus; arrow: mislocalized HDAC1. Scale bar: 50 μm. Right histogram: Quantification of γH2AX+ and HDAC1 mislocalized cells in the WT or 1-, 6-, and 12-month-old FTLD-TDP Tg mice. n = 4 sections per mouse, N = 5 mice per group, data are presented as mean ± SEM. ** p = 0.0079, **** p < 0.0001 by multiple comparison.
HDAC1 Dysregulation Induces Aberrant Cell Cycle and DNA Damage in Progress of TDP-43 Proteinopathies (2020)
Figure 5

Figure 5. TDP-43 interacts with HDAC1 and traps HDAC1 in inclusion bodies.

Left panel: flag-tagged full-length HDAC1 was overexpressed with myc-tagged TDP-43 in HEK-293T cells; the cell lysates were immunoprecipitated for flag and immunoblotted for TDP-43 and flag. Right panel: myc-tagged TDP-43 was overexpressed with flag-tagged full-length HDAC1 in HEK-293T cells; the cell lysates were immunoprecipitated for myc and immunoblotted for flag and TDP-43.

Upper left: flag-tagged full-length HDAC1 (b.Ⅰ) or various truncation mutations (b.Ⅱ-Ⅳ) were overexpressed with myc-tagged TDP-43; the catalytic domain is indicated in red. Lower panel: the western blotting of cell lysates immunoprecipitated for flag and immunoblotted for TDP-43.

Upper panel: Immunoprecipitation of cytosolic HDAC1 and immunoblotting of HDAC1 and TDP-43 in WT and FTLD-TDP Tg mice. Lower histogram: Quantification of immunoprecipitation results of HDAC1 and TDP-43 in WT and Tg mice. N = 5 mice per group, data are presented as mean ± SEM (%), * p =0.0149, *** p =0.0003 by t-test.

Western-blot of HDAC1 and TDP-43 in urea-soluble fractions. N = 5 mice per group.
HDAC1 Dysregulation Induces Aberrant Cell Cycle and DNA Damage in Progress of TDP-43 Proteinopathies (2020)
Figure 7

Figure 7. Pharmaceutical gain of HDAC1 activity repairs aberrant cell cycle activity, DNA damage, and neuronal loss.

Nuclear HDAC1 activity in mice after 2 months of compound 5104434 treatment. The nuclear extracts of the frontal cortices and hippocampus from 8-mon-old of mice were separated from total tissue lysates and were immunoprecipitated for HDAC1, and were then conducted to HDAC1 activity assay. N = 6 mice per group. TSA: nuclear extracts that were treated with Trichostatin A (TSA, an HDAC inhibitor) as a negative control for HDAC1-transferred fluorescent activity during the HDAC1 activity assay. * p = 0.0203 (WT vs Tg-Veh) or 0.0408 (Tg-Veh vs Tg+5104434) by multiple comparison.

Representative data of western blot for nuclear acetylated-histone 3 (Lys 9/14), total histone H3, and quantification of protein levels. N = 6 mice per group. * p = 0.0105 or *** p = 0.0008 by multiple comparison.

Representative data of western blot for nuclear HDAC1 levels and semi-quantification of the expression levels. N = 6 mice per group. ** p = 0.0022 (Tg)/ 0.0012 (Tg+5104434) by multiple comparison.

Left panel: representative data of western blot for cell cycle- and DNA damage-related proteins. Right histogram: semi-quantification of protein levels. N = 6 per group. *WT+Vehicle mice vs. Tg+Vehicle mice, #Tg+5104434 mice vs. Tg+Vehicle mice. Statistical analysis by multiple t-test with FDR correction, Q = 1%. **P ≤ 0.01; ###P ≤ 0.001 and ****/####P ≤ 0.0001. Exact p values are shown in Appendix Table S1.

IF staining of γH2AX in the frontal cortices and quantification of the immunoreactive cells from 8-mon-old of WT+Vehicle, Tg+Vehicle, Tg+5104434. n = 9 sections per mouse, N = 6 mice per group. Scale bar: 50 μm. * p = 0.0421 or **** p < 0.0001 by multiple comparison.

Immunohistochemistry of NeuN and the quantification of the immunoreactive cells in the hippocampus of three groups of mice, 8-mon-old WT+Vehicle, Tg+Vehicle and Tg+5104434. n = 9 sections per mouse, N = 6 mice per group. Scale bar: 50 μm.* p = 0.0414 or *** p = 0.0006 by multiple comparison.

IF staining of GFAP and the quantification of the immunoreactive cells in the hippocampus of three groups of mice, 12-month-old WT+Vehicle, Tg+Vehicle and Tg+5104434. n = 9 sections per mouse, N = 6 mice per group. Scale bar: 50 μm. ** p = 0.002 and **** p < 0.0001 by multiple comparison.

Data information: All data are presented as mean ± SEM (%) except for (E, G) where number of cells in each view is presented.