Journal: bioRxiv

Article Title: Early enhanced control of Plasmodium yoelii infection in IL-10–deficient mice is independent of IFN-γ, IL-12, and the humoral response

doi: 10.64898/2026.03.23.713659

Figure Lengend Snippet: WT, Il10 -/- , μMT , μMTIl10 -/- , Ifngr1 -/- , and Ifngr1 -/- Il10 -/- mice were infected i.p. with 10 5 P. yoelii 17X pRBCs. ( A ) Representative parasitemia curve of the primary infection. WT mice were i.p. injected with either 200 μg of Rat IgG, 500 μg anti-IFN-γ, 200 μg anti-IL10R, or both anti-IFN-γ and anti-IL-10R at day 1 p.i. and every 3 days after. WT mice served as controls. ( B ) Area-under-the-curve analysis included data for the entire curve in A. ( C ) Representative parasitemia curve of the primary infection. ( D ) Area-under-the-curve analysis included data for the entire curve in C. ( E ) Representative parasitemia curve of the primary infection. Il10 -/- mice were i.p. injected with either 500 μg of Rat IgG, 500 μg anti-IFN-γ, 500 μg anti-IL-12p75, or both anti-IFN-γ and anti-IL-12p75 at day 0, 3, 6, and 9 p.i. WT mice served as controls. ( F ) Area-under-the-curve analysis included data for the entire curve in E. ( G ) Representative parasitemia curve of the primary infection. μMTIl10 -/- mice were i.p. injected with either 500 μg Rat IgG or 500 μg anti-IFN-γ at day 0, 3, 6, and 9 p.i. WT, Il10 -/- , and μMT mice served as controls. (H ) Representative survival curve for the primary infection in G. Mice were euthanized when the parasite burden exceeded 50% or when prominent signs of morbidity were observed. ( B, D, F ) A non-parametric Kruskal-Wallis test with a post hoc Dunn’s multiple comparisons test or H ) a Mantel-Cox test determined significance. * p < 0.05, *** p < 0.001, **** p < 0.0001, ns = non-significant.

Article Snippet: To determine the necessity of IL-12 and/or IFN-γ without IL-10 during infection, Il10 -/- mice were i.p. administered either 500 μg of Rat IgG (Sigma-Aldrich), 500 μg of anti-IL-12p75 (clone R2-9A5, BioXcell), or 500 μg of anti-IFN-γ (clone XMG1.2, BioXcell) diluted in sterile PBS on day 0, 3, 6, 9, and 12 p.i.

Techniques: Infection, Injection

Journal: bioRxiv

Article Title: Expression of the human immunomodulatory protein, human B7-1 (CD80), accelerates neuroinflammation, synaptic loss, microvascular instability and lethality in a murine mode of Alzheimer’s Disease

doi: 10.64898/2025.12.30.697078

Figure Lengend Snippet: A) HEK 293 cells transiently transfected with constructs for either HUMAN, MOUSE, or CHIMERIC exon-swapped B7-1-GFP. Two days post transfection cells were titrated with increasing concentrations of mouse CTLA-4, CD28, and p75 NTR hIgG1 fc-fusion proteins and subsequently treated with anti-human A647 antibody. The percent binding was determined by flow cytometry gated as the percentage of GFP positive cells that were also Alexa647 positive (double positive). Note that mp75 NTR does not bind human B7-1 expressing cells within the range of concentrations tested. B) IgV domain exon swapping strategy to generate the h:mB7-1 KI mouse. C) Southern blot showing specific human DNA fragment in h:mB7-1 chimeric mice. D and E) Splenocytes were isolated from WT and h:mB71 chimera animals using established methods and stimulated with either LPS or PMA/Ionomycin for 24 hours. After stimulation, cells were collected and stained with cell-surface markers for different immune populations (αmCD45, αmCD3, αmCD19, αmCD11b, αmF4/80, αmCD11c) and human or mouse B7-1. D) Shows flow cytometry histogram plots for B7-1 expression in immune populations as indicated. T-cells were gated as (Singlets, Live, CD45+, CD3+, CD19-), B-cells were gated as (Singlets, Live, CD45+, CD3-, CD19+), macrophages were gated as (Singlets, Live, CD45+, CD3-, CD19-, CD11b+, F4/80+), and dendritic cells were gated as (Singlets, Live, CD45+, CD3-, CD19-, CD11b-, F4/80-, CD11c+). (E) Data shows the average fold-increase in B7-1 expression, compared to untreated populations, upon treatment with LPS or PMA/Ionomycin for wt (N=7) and h:mB7-1 (N= 8) mice.

Article Snippet: The following primary antibodies were used: beta amyloid polyclonal antibody (CT695) rabbit Invitrogen ref. 51-2700, 1.0 μg/mL o.n., CD31 (R&D Systems, AF3628) - 1 μg/mL, overnight (o.n.) incubation, hB7-1 (R&D Systems, AF140)-7.5 mg/ml, 2 day incubation (followed with secondary biotinylated and streptavidin conjugated Alexa Fluor®), Iba1 (NBP2-19019)-1:1000, Laminin (NB300-144)-1:250, o.n. incubation, mouse B7-1 (R&D Systems, AF740)-2.5 mg/mL, o.n, anti-p75 (R&D Systems, AF1157)-1 ug/mL, o.n, anti-PDGFRb (ThermoFisher, 14-1402-82)-1:200, 2 day incubation, GFAP (Sigma, G3893)-1:400, o.n., CD11b (Biolegend, 163702) (1:500).

Techniques: Transfection, Construct, Binding Assay, Flow Cytometry, Expressing, Southern Blot, Isolation, Staining

Journal: bioRxiv

Article Title: Expression of the human immunomodulatory protein, human B7-1 (CD80), accelerates neuroinflammation, synaptic loss, microvascular instability and lethality in a murine mode of Alzheimer’s Disease

doi: 10.64898/2025.12.30.697078

Figure Lengend Snippet: (A) Representative confocal images of wt/wt, h:mB71/wt, wt/CRND8 and h:mB71/CRND8 mice showing DAPI stained nuclei, CD11b+ cells, Aβ deposition and p75 NTR upregulation. (B) wt/CRND8 and h:mB71/CRND8 express a significative upregulation of CD11b+ cells/area compares to wt/wt and h:mB71/wt mice, however, no difference was observed between h:mB71/CRND8 and wt/CRND8 mice. (C) In the dSubiculum of h:mB71/CRND8 mice more p75 NTR protein is detected as compared to wt/CRND8 mice. Data shown are mean +/- SEM. In (B)*p=0.0102, in (C) ****p<0.0001. One-way ANOVA followed by post hoc Tukey test. n.s not significant. N=3. Scale bars: 10µm

Article Snippet: The following primary antibodies were used: beta amyloid polyclonal antibody (CT695) rabbit Invitrogen ref. 51-2700, 1.0 μg/mL o.n., CD31 (R&D Systems, AF3628) - 1 μg/mL, overnight (o.n.) incubation, hB7-1 (R&D Systems, AF140)-7.5 mg/ml, 2 day incubation (followed with secondary biotinylated and streptavidin conjugated Alexa Fluor®), Iba1 (NBP2-19019)-1:1000, Laminin (NB300-144)-1:250, o.n. incubation, mouse B7-1 (R&D Systems, AF740)-2.5 mg/mL, o.n, anti-p75 (R&D Systems, AF1157)-1 ug/mL, o.n, anti-PDGFRb (ThermoFisher, 14-1402-82)-1:200, 2 day incubation, GFAP (Sigma, G3893)-1:400, o.n., CD11b (Biolegend, 163702) (1:500).

Techniques: Staining

Journal: bioRxiv

Article Title: Expression of the human immunomodulatory protein, human B7-1 (CD80), accelerates neuroinflammation, synaptic loss, microvascular instability and lethality in a murine mode of Alzheimer’s Disease

doi: 10.64898/2025.12.30.697078

Figure Lengend Snippet: (A) Representative confocal images of wt/wt, h:mB71/wt, wt/CRND8 and h:mB71/CRND8 mice showing DAPI stained nuclei, CD11b+ cells, Aβ deposition and p75 NTR . (B) At 4mo, in the dSubiculum of the h:B71/CRND8 mice, there is significant upregulation of CD11b+ cells/area compared to wt/CRND8 (p=values), wt/wt and h:mB71/wt mice (p=values). Most of the CD11b+ cells are near beta amyloid accumulation. (C) In the dSubiculum of h:mB71/CRND8 mice there is more p75 NTR protein detected than in wt/CRND8 mice, however p75 is detected near beta amyloid accumulation in both genotypes. Data shown are mean +/- SEM. In (B)****p<0.0001, ***p=0.0246. In (C) ****p<0.0001; ***p=0.0003 (between h:mB71 and CRND8) and ***p=0.0002 (WT and CRND8). One-way ANOVA followed by post hoc Tukey test). N=3. Scale bars: 10µm

Article Snippet: The following primary antibodies were used: beta amyloid polyclonal antibody (CT695) rabbit Invitrogen ref. 51-2700, 1.0 μg/mL o.n., CD31 (R&D Systems, AF3628) - 1 μg/mL, overnight (o.n.) incubation, hB7-1 (R&D Systems, AF140)-7.5 mg/ml, 2 day incubation (followed with secondary biotinylated and streptavidin conjugated Alexa Fluor®), Iba1 (NBP2-19019)-1:1000, Laminin (NB300-144)-1:250, o.n. incubation, mouse B7-1 (R&D Systems, AF740)-2.5 mg/mL, o.n, anti-p75 (R&D Systems, AF1157)-1 ug/mL, o.n, anti-PDGFRb (ThermoFisher, 14-1402-82)-1:200, 2 day incubation, GFAP (Sigma, G3893)-1:400, o.n., CD11b (Biolegend, 163702) (1:500).

Techniques: Staining

Journal: bioRxiv

Article Title: Expression of the human immunomodulatory protein, human B7-1 (CD80), accelerates neuroinflammation, synaptic loss, microvascular instability and lethality in a murine mode of Alzheimer’s Disease

doi: 10.64898/2025.12.30.697078

Figure Lengend Snippet: Representative confocal images of wt/wt, h:mB71/wt, wt/CRND8 and h:mB71/CRND8 mice showing DAPI stained nuclei, GFAP+ cells, VE-CAD and p75 NTR protein. Scale bars: 10 µm

Article Snippet: The following primary antibodies were used: beta amyloid polyclonal antibody (CT695) rabbit Invitrogen ref. 51-2700, 1.0 μg/mL o.n., CD31 (R&D Systems, AF3628) - 1 μg/mL, overnight (o.n.) incubation, hB7-1 (R&D Systems, AF140)-7.5 mg/ml, 2 day incubation (followed with secondary biotinylated and streptavidin conjugated Alexa Fluor®), Iba1 (NBP2-19019)-1:1000, Laminin (NB300-144)-1:250, o.n. incubation, mouse B7-1 (R&D Systems, AF740)-2.5 mg/mL, o.n, anti-p75 (R&D Systems, AF1157)-1 ug/mL, o.n, anti-PDGFRb (ThermoFisher, 14-1402-82)-1:200, 2 day incubation, GFAP (Sigma, G3893)-1:400, o.n., CD11b (Biolegend, 163702) (1:500).

Techniques: Staining

Journal: Scientific Reports

Article Title: Disease-specific alterations of the enteric nervous system in precancerous colonic mucosa and their implications for mucin regulation

doi: 10.1038/s41598-025-24480-z

Figure Lengend Snippet: Spatiotemporal distribution of mucosal enteric glial cells (EGCs) in the human ileum and colon. ( A ) Representative images of mucosal EGCs in the adult human colon. Sox10 + (brown) cells with p75NTR- or CD56-positive fine processes (red) in the human colonic mucosa (arrowhead). Scale bar: 20 μm. ( B ) Mouse mucosal EGCs (green) targeted in the mGFAP-Cre-GFP reporter model. Scale bar: 20 μm. ( C ) Representative images of mucosal Sox10 + cells in the distal ileum, the right-sided colon (R/C), and the left-sided colon (L/C). Sox10 + cells are widely distributed in the lamina propria, with higher concentrations around the crypt base (arrow head). Lower panels show the number of mucosal Sox10 + cells in the ileum, R/C, and L/C of individuals of different ages and sexes. Scale bar: 100 μm. ( D ) Multivariate multiple regression analysis shows that the number of Sox10 + cells is lower in the ileum compared to the R/C and L/C (left), and lower in females than in males (middle). The proportion of Sox10 + cells in the luminal part of the mucosa is lower in the older group (right). * p < 0.05, *** p < 0.01.

Article Snippet: Cells were incubated with either rabbit anti-Sox10 (1/200 dilution, EP268, Cell Marque, Rocklin, CA) or mouse anti-p75NTR antibody (1/100 dilution, ME20.4, Santa Cruz Biotechnology) at 4 °C overnight followed by visualization with the Alexa Fluor 488-conjugated secondary antibodies.

Techniques:

Journal: iScience

Article Title: Cholinotrophic basal forebrain connectome dysfunction in Down syndrome with and without dementia

doi: 10.1016/j.isci.2025.113041

Figure Lengend Snippet: Images and quantitation of cholinotrophic basal forebrain neuron pathology within the nucleus basalis of Meynert in DSD+, DSD- and AMC cases (A–N) Photomicrographs show p75 NTR (brown, A-C), ChAT (purple, D-F) positive cells in AMC, DSD-, and DSD+ cases. Note the decrease in the number and intensity of both p75 NTR and ChAT labeled neurons in DSD- cases compared to an even greater reduction in DSD+ and globose shaped cells (black arrows) in DSD+ cases (C, F). Arrows in (A–F) mark cells shown at a higher magnification in the boxed areas located at the lower right corner of each panel. Dark brown AT8 (G, I) and TauC3 (H, J) bearing neurofibrillary tangles (NFTs) were observed only in the DSD- and DSD+ cases. Black arrows indicate globose shaped NFTs (G, H, and I) that are shown at a higher magnification in boxed areas adjacent to the lower magnification images. Note that not all neurons within the nbM (thin black arrows) contained tau pathology (G, H) in DSD- cases. Moreover, TauC3 staining revealed two NFT phenotypes that displayed either peripherally located or intense labeling that filled the entire structure (see boxed images adjacent to (H) and (J)). Scale bar in F = 50 μm and inset = 20 μm applies to panels A-E. Scale bar in J = 20 μm and inset = 20 μm applies to (G–J). Histograms show a significant reduction in both p75 NTR (K) and ChAT (L) positive cells in DSD+ compared to AMC. Although no significant were found in number of AT8 or TauC3 NFT positive cells (M), there was an increase in NTs in DSD+ compared to DSD- (N). ACM n = 5, DSD- n = 5, DSD+ n = 10. Data shown are presented as mean ± SEM. Statistical significance was determined using the Kruskal–Wallis’s test followed by Dunn’s test for comparisons across three clinical groups, and the Mann–Whitney test for comparisons between two groups (DSD- vs. DSD+). Significance levels (∗) were set at: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Antibodies were added to blocking buffer and membranes incubated overnight (4°C) in mouse anti-p75 NTR ([1:150], Santa Cruz Biotechnology Cat# sc-271708, RRID: AB_10714958 ), rabbit anti-ChAT ([1:500] (Proteintech Cat# 20747-1-AP, RRID: AB_10898169 ), rabbit anti-TrkA ([1:150], Fitzgerald Industries International Cat# 20R-TR013, RRID: AB_1289098 ) or rabbit anti-proNGF ([1:250], Alomone Labs Cat# ANT-005, RRID: AB_2040021 ), washed, incubated with horseradish peroxidase-conjugated with either a goat anti-mouse IgG (1:200) or goat anti-rabbit IgG (1:200) secondary antibody at RT, visualized by chemiluminescence (Kodak Image Station 440CF; Perkin-Elmer, Wellesley, MA), and quantified with Kodak 1.

Techniques: Quantitation Assay, Labeling, Staining, MANN-WHITNEY

Journal: iScience

Article Title: Cholinotrophic basal forebrain connectome dysfunction in Down syndrome with and without dementia

doi: 10.1016/j.isci.2025.113041

Figure Lengend Snippet: Immunofluorescent images and quantification of p75 NTR neurons with and without AT8 and MAP2, as well as Thioflavin S single and AT8/Thioflavin S dual-labeled nbM cells in DSD+ and DSD-cases. (A–U) Immunofluorescent neurons labeled with p75 NTR (A, E, green), MAP2 (B, F, red), AT8 (C, G, Cyan), ThS (blue) (I, L) and merge images (D, H, K, N, R) in DSD- and DSD+ cases. In the DSD-cases there were greater p75 NTR cells AT8 negative neurons and p75 NTR MAP2 dual labeled neurons (A-D) compared to the DSD+ cases (E–H). Note that not all p75 NTR cells colocalize with AT8 or MAP2 in both DS groups (D, H). To determine the stage of a tangle tissue was stained for ThS, a marker of advanced pathology and the early stage AT8 phosphorylation antibody. Note that there are only a few ThS-labeled tangles that displayed AT8 in both DS groups (yellow arrows), compared to single ThS tangles, which were greater in DSD+ than in DSD- (white arrows) (I-L). Note that ThS positive NFTs that do not contain AT8 also do not colocalize with MAP2 (O-R). Scale bar in F = 25 μm applies to panels A-G, N = 25 μm applies to panels I-M and R = 10 μm and applies to O-Q. Graph showing a significant reduction in both p75 NTR AT8 immuno-negative, and p75 NTR MAP2 dual labeled neurons in DSD+ compared to DSD- (S). ThS-positive cells were greater (T), while the percentage of double-labeled cells with AT8 and ThS decreased (U) in DSD+ compared to individuals without dementia. DSD- n = 5, DSD+ n = 5. Data are presented as mean ± SEM. Statistical significance was determined using Mann–Whitney test for comparisons between DSD- and DSD+. Significance levels (∗) were set at: ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Antibodies were added to blocking buffer and membranes incubated overnight (4°C) in mouse anti-p75 NTR ([1:150], Santa Cruz Biotechnology Cat# sc-271708, RRID: AB_10714958 ), rabbit anti-ChAT ([1:500] (Proteintech Cat# 20747-1-AP, RRID: AB_10898169 ), rabbit anti-TrkA ([1:150], Fitzgerald Industries International Cat# 20R-TR013, RRID: AB_1289098 ) or rabbit anti-proNGF ([1:250], Alomone Labs Cat# ANT-005, RRID: AB_2040021 ), washed, incubated with horseradish peroxidase-conjugated with either a goat anti-mouse IgG (1:200) or goat anti-rabbit IgG (1:200) secondary antibody at RT, visualized by chemiluminescence (Kodak Image Station 440CF; Perkin-Elmer, Wellesley, MA), and quantified with Kodak 1.

Techniques: Labeling, Staining, Marker, Phospho-proteomics, MANN-WHITNEY

Journal: iScience

Article Title: Cholinotrophic basal forebrain connectome dysfunction in Down syndrome with and without dementia

doi: 10.1016/j.isci.2025.113041

Figure Lengend Snippet: FC cholinotrophic protein levels in AMC, DSD- and DSD+ (A–D) Representative immunoblots, and bar graphs show a significant upregulation of (A) proNGF and (B) p75 NTR , while (C) ChAT protein was downregulated between AMC and DSD+. (D) TrkA protein levels were stable across the groups analyzed. ACM n = 5, DSD- n = 5, DSD+ n = 13. Data are presented as mean ± SEM. Statistical significance was determined using the Kruskal–Wallis’s test followed by Dunn’s test for comparisons across three clinical groups. Significance levels (∗) were set at: ∗ p < 0.05.

Article Snippet: Antibodies were added to blocking buffer and membranes incubated overnight (4°C) in mouse anti-p75 NTR ([1:150], Santa Cruz Biotechnology Cat# sc-271708, RRID: AB_10714958 ), rabbit anti-ChAT ([1:500] (Proteintech Cat# 20747-1-AP, RRID: AB_10898169 ), rabbit anti-TrkA ([1:150], Fitzgerald Industries International Cat# 20R-TR013, RRID: AB_1289098 ) or rabbit anti-proNGF ([1:250], Alomone Labs Cat# ANT-005, RRID: AB_2040021 ), washed, incubated with horseradish peroxidase-conjugated with either a goat anti-mouse IgG (1:200) or goat anti-rabbit IgG (1:200) secondary antibody at RT, visualized by chemiluminescence (Kodak Image Station 440CF; Perkin-Elmer, Wellesley, MA), and quantified with Kodak 1.

Techniques: Western Blot

Journal: iScience

Article Title: Cholinotrophic basal forebrain connectome dysfunction in Down syndrome with and without dementia

doi: 10.1016/j.isci.2025.113041

Figure Lengend Snippet: Summary of changes in the cholinotrophic basal forebrain connectome in DS Diagrammatic sagittal view of the human brain (A) and a modified stacked bar graph (B) illustrating differences in the pathobiology of the cholinotrophic projection system between non-trisomy age-matched control (AMC), DS without dementia (DSD-) and DS with dementia (DSD+) individuals with DS. Frontal cortex protein levels for ChAT (pink), proNGF (green), p75 NTR (purple) and TrkA (light blue). Nucleus basalis NFTs of ThS (blue), AT8 (orange) and TauC3 (black) and counts of p75 NTR and ChAT (red) neurons. Created with BioRender.com .

Article Snippet: Antibodies were added to blocking buffer and membranes incubated overnight (4°C) in mouse anti-p75 NTR ([1:150], Santa Cruz Biotechnology Cat# sc-271708, RRID: AB_10714958 ), rabbit anti-ChAT ([1:500] (Proteintech Cat# 20747-1-AP, RRID: AB_10898169 ), rabbit anti-TrkA ([1:150], Fitzgerald Industries International Cat# 20R-TR013, RRID: AB_1289098 ) or rabbit anti-proNGF ([1:250], Alomone Labs Cat# ANT-005, RRID: AB_2040021 ), washed, incubated with horseradish peroxidase-conjugated with either a goat anti-mouse IgG (1:200) or goat anti-rabbit IgG (1:200) secondary antibody at RT, visualized by chemiluminescence (Kodak Image Station 440CF; Perkin-Elmer, Wellesley, MA), and quantified with Kodak 1.

Techniques: Modification, Control