Journal: Scientific Reports

Article Title: Single neonatal dexamethasone administration has long-lasting outcome on depressive-like behaviour, Bdnf, Nt-3, p75ngfr and sorting receptors ( SorCS1-3 ) stress reactive expression

doi: 10.1038/s41598-021-87652-7

Figure Lengend Snippet: P75NTR protein level changes in the brainstem after single DEX injection 0.2 mg/kg to P2 rat pups. ( A ) p75NTR protein level timeline profile ONE WAY ANOVA, * p < 0.05 compared to the control group, Data are presented as M ± SE ( B ) representative western blot image, (original images are in ), ( C ) Immunohistochemistry results.Mean p75NTR fluorescence intensity in norepinephrine LC neurons. ONE WAY ANOVA, * p < 0.05 compared to the control group, Data are presented as M ± SE ( D ) representative micro-photographs of p75NTR (green, Alexa 488) protein level changes in LC norepinephrine neurons(red, Alexa 568) after DEX administration. P75NTR—green (Alexa 488), Tyrosine Hydroxylase (TH)—red (Alexa 568), scale bar—100 mm.

Article Snippet: Subsequently section were incubated overnight at 4°C with primary antibodies: AB1542 sheep anti tyrosine hydroxylase, Millipore 1:300, 4201 monoclonal rabbit anti p75NTR (D4B3), Cell Signaling 1:300 diluted with blocking buffer.

Techniques: Injection, Control, Western Blot, Immunohistochemistry, Fluorescence

Journal: iScience

Article Title: Muscle denervation promotes functional interactions between glial and mesenchymal cells through NGFR and NGF

doi: 10.1016/j.isci.2023.107114

Figure Lengend Snippet: Chromatin accessibility at single nucleus level of muscle-resident cells upon denervation (A) UMAP embedding of snATAC-seq data after integration, colored according to our scRNA-seq data. (B) Table illustrating the number of nuclei assigned to each muscle-resident cell population and the number of differential accessibility regions (DARs) between DEN5d and CTR. (C) Histogram of the number of DARs overlapping promoters (red, UP) or non-promoter (enhancer, in blue, DOWN) regions, per cell type. (D–F) UMAP embedding of snATAC-seq data, showing the predicted gene expression profile for Thy1 (CD90) (D), Ngf (E) ,and Ngfr (F), either dim (blue) or bright (red).

Article Snippet: Anti-p75NTR (D4B3) XP®Rabbit antibody , Cell Signaling Technology , Cat# 8238; RRID: AB_10839265.

Techniques: Gene Expression

Journal: iScience

Article Title: Muscle denervation promotes functional interactions between glial and mesenchymal cells through NGFR and NGF

doi: 10.1016/j.isci.2023.107114

Figure Lengend Snippet: Ligand-receptor interaction analysis (A) Heatmap plot from CellPhoneDB showing the total number of interactions (log count) between cell types in our scRNA-seq data. Interaction log count visualized on a scale from low (blue) to high (dark red). (B) Venn diagram of the intersection between molecular pathways downstream to receptors on glial cells predicted to interact with ligands expressed by activated fibroblasts (Predicted), and molecular pathways predicted to be activated in glial cells based on gene expression (Activated). (C) Heatmap containing the molecular pathways connected to a specific receptor-ligand pair (only downstream of NGF-NGFR; complete information can be found in Figure S9 ) for the interactions between glial cells and activated fibroblasts (Interactome). On the right each pathway activation status calculated by Ingenuity Pathway Analysis (IPA; red for activation, blue for repression, and gray or white for activation status info missing) at DEN2d, DEN5d, and DEN15d, respectively compared to the control. EMT: epithelial-mesenchymal transition; NO: nitric oxide; ROS: reactive oxygen species. Pathways activation status (Z-scores) visualized on a scale from repressed (blue) to activated (red), with neutral status (Z-Score = 0) set to white. Pathways not reported in a specific timepoint, with Z-score = N/A, are represented in grey. (D) Representative images of CD90 (cyan) and NGF (green) immunofluorescence staining of TA sections from Control (CTR, upper) and denervated (DEN5d, lower) Plp1-TdTomato (red) mice. Nuclei were counterstained with DAPI (white). Serial section was stained for Caveolin-3 (magenta). Scale bar: 100μm. N = 4 mice. (E) Representative z stack images of whole mount staining of NGFR (yellow) and NGF (red) in extensor digitorum longus (EDL) isolated from CTR and DEN5d B6 mice. Acetylcholine receptor (AChR) at post-synaptic membrane of myofiber was stained with αBTX (green): α-Bungarotoxin. Nuclei were counterstained with Hoechst (blue). Scale bar: 50μm. N = 3 mice.

Article Snippet: Anti-p75NTR (D4B3) XP®Rabbit antibody , Cell Signaling Technology , Cat# 8238; RRID: AB_10839265.

Techniques: Gene Expression, Activation Assay, Control, Immunofluorescence, Staining, Isolation, Membrane

Journal: iScience

Article Title: Muscle denervation promotes functional interactions between glial and mesenchymal cells through NGFR and NGF

doi: 10.1016/j.isci.2023.107114

Figure Lengend Snippet: Recombinant NGF promotes expansion of NGFR-positive glail cells ex vivo (A) FACS plot showing the sorting strategy for CD90 pos SCA1 pos activated fibroblasts (abbreviated as CD90 pos cells), CD59A pos NGFR neg cells (abbreviated as CD59A pos cells), and CD59A pos NGFR pos Glial cells (abbreviated as NGFR pos cells) simultaneously from control (upper, CTR) and DEN5d (bottom) muscles. (B) Percentage of 3 populations (gated in A) by FACS at CTR and DEN5d conditions. N = 8 mice. (C) Sorted NGFR pos glial cells were treated with 0/5/10/20 ng/mL beta-NGF for 2 days in culture. Hoechst and propidium iodide (PI) were added into the medium to help detect total cell number and dead cell number, respectively. Images were captured at day 0 and day 2. Quantified live cell number at day 2 was normalized to that at day 0. N = 5 mice. In B and C, data are represented as mean ± SD. Statistical significance was analyzed by two-way ANOVA in (B) and one-way ANOVA in (C) with p values shown as ∗p < 0.05 and ns for not significant.

Article Snippet: Anti-p75NTR (D4B3) XP®Rabbit antibody , Cell Signaling Technology , Cat# 8238; RRID: AB_10839265.

Techniques: Recombinant, Ex Vivo, Control, Muscles

Journal: iScience

Article Title: Muscle denervation promotes functional interactions between glial and mesenchymal cells through NGFR and NGF

doi: 10.1016/j.isci.2023.107114

Figure Lengend Snippet: Co-culture with NGF-expressing mesenchymal cells promotes expansion of NGFR-positive glial cells ex vivo (A) Graphic illustration of co-culture experiment with NGFR pos cells and CTR- or DEN5d- CD90 pos cells (abbreviated as CTR group or DEN5d group, respectively). Cells were fixed after 3 days culture for EdU assay (EdU was added for 4 h pulsing before fixation) and TUNEL assay, respectively. (B) Representative cell staining images of SOX10 (green) and EdU (magenta), counterstained with Hoechst (blue). Arrow heads indicate SOX10 pos EdU pos Glial cells. Scale bar: 100μm. (C) Representative cell staining images of SOX10 (green) and TUNEL (red), counterstained with Hoechst (blue). Arrows indicate SOX10 pos TUNEL pos Glial cells. Scale bar: 100μm. (D) SOX10 pos Glial cell number per field was quantified in two co-culture groups. By normalization to the CTR group, relative Glial cell number was shown. N = 6 mice. (E) EdU pos percentage of SOX10 pos Glial cells in two co-culture groups. N = 6 mice. (F) TUNEL pos percentage of SOX10 pos Glial cells in two co-culture groups. N = 3 mice. Statistical significance was analyzed by paired Student’s t test in (D-F) with p values shown as ∗∗p < 0.01 and ns for not significant.

Article Snippet: Anti-p75NTR (D4B3) XP®Rabbit antibody , Cell Signaling Technology , Cat# 8238; RRID: AB_10839265.

Techniques: Co-Culture Assay, Expressing, Ex Vivo, EdU Assay, TUNEL Assay, Staining

Journal: iScience

Article Title: Muscle denervation promotes functional interactions between glial and mesenchymal cells through NGFR and NGF

doi: 10.1016/j.isci.2023.107114

Figure Lengend Snippet:

Article Snippet: Anti-p75NTR (D4B3) XP®Rabbit antibody , Cell Signaling Technology , Cat# 8238; RRID: AB_10839265.

Techniques: Recombinant, Electron Microscopy, Imaging, TUNEL Assay, In Situ, Reverse Transcription, SYBR Green Assay, Software, Microscopy

Journal: Scientific Reports

Article Title: Single neonatal dexamethasone administration has long-lasting outcome on depressive-like behaviour, Bdnf, Nt-3, p75ngfr and sorting receptors ( SorCS1-3 ) stress reactive expression

doi: 10.1038/s41598-021-87652-7

Figure Lengend Snippet: P75NTR protein level changes in the brainstem after single DEX injection 0.2 mg/kg to P2 rat pups. ( A ) p75NTR protein level timeline profile ONE WAY ANOVA, * p < 0.05 compared to the control group, Data are presented as M ± SE ( B ) representative western blot image, (original images are in ), ( C ) Immunohistochemistry results.Mean p75NTR fluorescence intensity in norepinephrine LC neurons. ONE WAY ANOVA, * p < 0.05 compared to the control group, Data are presented as M ± SE ( D ) representative micro-photographs of p75NTR (green, Alexa 488) protein level changes in LC norepinephrine neurons(red, Alexa 568) after DEX administration. P75NTR—green (Alexa 488), Tyrosine Hydroxylase (TH)—red (Alexa 568), scale bar—100 mm.

Article Snippet: Subsequently section were incubated overnight at 4°C with primary antibodies: AB1542 sheep anti tyrosine hydroxylase, Millipore 1:300, 4201 monoclonal rabbit anti p75NTR (D4B3), Cell Signaling 1:300 diluted with blocking buffer.

Techniques: Injection, Control, Western Blot, Immunohistochemistry, Fluorescence

Journal: Cell and Tissue Research

Article Title: P75 neurotrophin receptor controls subventricular zone neural stem cell migration after stroke

doi: 10.1007/s00441-021-03539-z

Figure Lengend Snippet: p75 NTR −/− SVZ NSPCs reveal and altered morphology and cytoskeletal organization. a Immunoblot protein expression of ArhGap5 in p75 NTR−/− compared to WT NSPCs. b Quantification graph of Arhgap5 protein expression in p75 NTR−/− compared to WT NSPCs. c – h Immunolabeling for phalloidin (green, left) and α-tubulin (gray, middle and enlargement right) of BMP-2 treated p75 NTR−/− and WT neurospheres in vitro. i Quantification graph of α-tubulin immunoreactivity in p75 NTR−/− compared to WT neurospheres. Scale bars = 35 µm ( c , d , f , g ), 20 µm ( e – h ). Values are mean ± SEM ( p values calculated Student’s t test; * p < 0.05, ** p < 0.01. Representative Western blot is shown, n = 4

Article Snippet: The following primary antibodies were used: rabbit anti-p75 NTR (1:2000, Cell Signaling, D4B3), P-Smad1/5/8 (1:1000, Cell Signaling), rabbit anti-Smad1 (1:1000, Cell Signaling), rabbit anti-GAPDH (1:1000, Cell Signaling), rabbit ArhGap5 (1:2000, Cell Signaling), and rabbit anti-BDNF (1:1000, Abcam, #ab108319).

Techniques: Western Blot, Expressing, Immunolabeling, In Vitro