probdnf  (Alomone Labs)

Journal: Frontiers in Aging Neuroscience

Article Title: Increased Hippocampal ProBDNF Contributes to Memory Impairments in Aged Mice

doi: 10.3389/fnagi.2017.00284

Figure Lengend Snippet: Memory impairment in aged mice positively correlates with increases in hippocampal proBDNF levels. (A) Average (±SEM) reference memory (RM) and working memory (WM) errors over four 4-session blocks of a WRAM task in 24-month old aged mice and 4-month old young mice. RM and WM errors positively correlate with proBDNF levels. (B) Increase in proBDNF, but not brain-derived neurotrophic factor (BDNF), in 24-month old aged mice relative to young 4-month old mice. (C) Relative to young 4-month old mice, aged 24-month old mice show increased p75 Neurotrophin Receptor (p75NTR) and decreased p-trk140 and trk140. (D) Relative to young 4-month old mice, aged 24-month old mice show decreased Tissue Plasminogen Activator (tPA), but not carboxypeptidase E (CPE). * p

Article Snippet: On d20–22 unimpaired mice in group “unimpaired + proBDNF” were injected with “uncleavable ” mouse proBDNF (proBDNF mut-mouse, Alomone, Jerusalem, Israel) 40 pg/0.4 μL/side.

Techniques: Mouse Assay, Derivative Assay

Journal: Frontiers in Aging Neuroscience

Article Title: Increased Hippocampal ProBDNF Contributes to Memory Impairments in Aged Mice

doi: 10.3389/fnagi.2017.00284

Figure Lengend Snippet: Theoretical model of the role of proBDNF in learning in memory in aged individuals. In young individuals, maturation of proBDNF to BDNF is controlled by plasmin and tPA, and is essential for learning and memory. Aged individuals show decreased levels of tPA and plasmin, and increased levels of uncleaved proBDNF, associated with increased spine remodeling and memory deficits. Blockade of p75NTR (e.g., by TAT-Pep5, as in the current study) leads to spine growth, and rescues learning and memory.

Article Snippet: On d20–22 unimpaired mice in group “unimpaired + proBDNF” were injected with “uncleavable ” mouse proBDNF (proBDNF mut-mouse, Alomone, Jerusalem, Israel) 40 pg/0.4 μL/side.

Techniques:

Journal: Frontiers in Aging Neuroscience

Article Title: Increased Hippocampal ProBDNF Contributes to Memory Impairments in Aged Mice

doi: 10.3389/fnagi.2017.00284

Figure Lengend Snippet: Timeline of Experiments 1 and 2. In Experiment 2, mice were split into three groups based on their performance (unimpaired/impaired) and henceforth infused with different drugs (saline, proBDNF, TAT-Pep5). d, day of study; WRAM, behavioral training/testing in the water radial arm maze task; SAL, saline.

Article Snippet: On d20–22 unimpaired mice in group “unimpaired + proBDNF” were injected with “uncleavable ” mouse proBDNF (proBDNF mut-mouse, Alomone, Jerusalem, Israel) 40 pg/0.4 μL/side.

Techniques: Mouse Assay

Journal: Frontiers in Aging Neuroscience

Article Title: Increased Hippocampal ProBDNF Contributes to Memory Impairments in Aged Mice

doi: 10.3389/fnagi.2017.00284

Figure Lengend Snippet: Intra-hippocampal infusion of proBDNF impairs memory in well-performing (unimpaired) mice, while TAT-Pep5 infusion improves memory in poorly-performing (impaired) mice. (A) Average (±SEM) RM and WM errors in memory impaired 18-month old mice receiving intra-hippocampal saline infusions (impaired + SAL, n = 8, open triangles) and well-performing 18-month old aged mice receiving uncleavable proBDNF intra-hippocampal infusions (unimpaired + proBDNF, n = 16, open circles) over four daily sessions of a WRAM task. (B) Average (± SEM) RM and WM errors in memory impaired 18-month old mice infused with saline (impaired + SAL, n = 8, open triangles) and memory impaired 18-month old mice receiving intra-hippocampal infusions of TAT-Pep5 (impaired + TAT-Pep5, n = 9, closed circles) over four daily sessions of a WRAM task. (C) Representative images indicating the locations of drug infusions at two levels of the hippocampus. ns not significant; * p

Article Snippet: On d20–22 unimpaired mice in group “unimpaired + proBDNF” were injected with “uncleavable ” mouse proBDNF (proBDNF mut-mouse, Alomone, Jerusalem, Israel) 40 pg/0.4 μL/side.

Techniques: Mouse Assay

Journal: Frontiers in Aging Neuroscience

Article Title: Increased Hippocampal ProBDNF Contributes to Memory Impairments in Aged Mice

doi: 10.3389/fnagi.2017.00284

Figure Lengend Snippet: Intra-hippocampal infusion of proBDNF increases p-cofilin levels in memory-unimpaired mice to levels seen in memory-impaired mice. (A) p-Cofilin to total cofilin ratio in memory-unimpaired mice, memory-impaired mice, and memory-unimpaired mice infused with proBDNF. (B) Representative p-cofilin and cofilin blots. ** p

Article Snippet: On d20–22 unimpaired mice in group “unimpaired + proBDNF” were injected with “uncleavable ” mouse proBDNF (proBDNF mut-mouse, Alomone, Jerusalem, Israel) 40 pg/0.4 μL/side.

Techniques: Mouse Assay

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Schematic diagram showing how proBDNF dampens CD4 + T cell activity and contributes to the pathogenesis of SAE. In sepsis, proBDNF expression is increased in peripheral blood and meningeal immune cells, which then decreases the infiltration of CD4 + T cells in the meninges. As a result, meningeal pro-inflammatory cytokines such as IL-6 and IL-1β are upregulated, but anti-inflammatory cytokines including IL-4 and IL-13 are downregulated, finally leading to SAE. SAE, sepsis-associated encephalopathy

Article Snippet: As shown, proBDNF protein did not affect T cell subgroups in normal mice (Fig. a–c).

Techniques: Activity Assay, Expressing

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Exogenous proBDNF protein reduced CD4 + T cells but increased CD8 + T cells of septic mice in vitro. Mice injected with saline or LPS (5 mg kg −1 ) for 5 days and the splenocytes were isolated and cultured for 3 days in vitro. Exogenous proBDNF did not alter the percentage of a CD3 + T cells in CD45 + cells or the percentage of b CD4 + T cells or c CD8 + T cells in CD3 + T cells in splenocytes from mice treated with saline. d Exogenous proBDNF did not alter the percentage of CD3 + T cells in CD45 + cells in splenocytes in septic mice. e–f ProBDNF treatment significantly decreased the percentage of e CD4 + T cells but increased the percentage of f CD8 + T cells in CD3 + T cells in splenocytes in LPS-treated mice. n = 4 in each group. Data were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

Article Snippet: As shown, proBDNF protein did not affect T cell subgroups in normal mice (Fig. a–c).

Techniques: Mouse Assay, In Vitro, Injection, Isolation, Cell Culture

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Increased proBDNF expression in meningeal and peripheral blood immune cells in septic mice. Mice were i.p. injected with LPS (5 mg kg −1 ) and meninges and peripheral blood were harvested for immunofluorescence staining or flow cytometry. a Representative whole mount meningeal immunofluorescence images showed markedly increased proBDNF-positive staining cells in the meninges in mice at 1 day after LPS injection compared to saline injected mice. The high magnification images around the arrows are displayed in a white square as insets. Bar = 100 μm. b – f Representative meningeal single cell flow cytometry images ( upper panel ) and its statistical analysis ( lower panel ) indicated that proBDNF MFI was increased in meningeal b CD3 + T cells, c CD4 + T cells, d CD8 + T cells, and f CD11b + monocytes/macrophages at 1 day after LPS injection. proBDNF in meningeal e CD19 + B cells upregulated until 5 days after LPS injection. n = 10 in the Con group, n = 6 in LPS groups. g – k Upregulation of proBDNF in g CD3 + T cells, h CD4 + T cells, i CD8 + T cells, j CD19 + B cells, and k CD11b + monocytes/macrophages in peripheral blood in LPS-injected mice were detected. n = 9 in Con group, n = 8 in the LPS1d group, n = 4 in the LPS 5d group. Data b – k were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

Article Snippet: As shown, proBDNF protein did not affect T cell subgroups in normal mice (Fig. a–c).

Techniques: Expressing, Mouse Assay, Injection, Immunofluorescence, Staining, Flow Cytometry

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Effect of i.c.v. injection of anti-proBDNF antibody on fear memory and meningeal immune activity in the septic mice. Mice were bilateral i.c.v. injected with 1 μg McAb-proB 3 days before LPS injection. Behavior tests were performed 1 day after LPS injection. a Representative images showed the broad and thorough diffusion of drugs in cerebroventricular lumen following i.c.v. injection with methylene blue. b McAb-proB i.c.v. injection did not influence weight of mice. c–e There was no difference of c fear conditioning acquiring performance in each group, nor McAb-proB increased the freezing time of d contextual or e cued fear conditioning test as compared to IgG control after LPS injection. n = 6 in each group. Data b , c , and e were analyzed by repeated measures ANOVA and followed by Bonferroni post hoc test and data d was analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

Article Snippet: As shown, proBDNF protein did not affect T cell subgroups in normal mice (Fig. a–c).

Techniques: Injection, Activity Assay, Mouse Assay, Diffusion-based Assay

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Systemic blockade of proBDNF ameliorated cognitive dysfunction and restored meningeal and peripheral CD4 + T cell ratio in septic mice. Mice were i.p. injected with proBDNF 30 min before LPS (5 mg kg −1 ) injection. Fear conditioning testing was performed 1 day after LPS injection. Meninges and peripheral blood were harvested 5 days after LPS injection for flow cytometry. a McAb-proB did not influence the weight of mice or b fear conditioning acquiring. c , d McAb-proB greatly alleviated memory deficit induced by LPS injection in mice as indicated by the increased freezing time in ( c ) contextual and ( d ) cued fear conditioning tests in the McAb-proB group relative to the IgG control. n = 8 in each group. Data a , b , and d were analyzed by repeated measures ANOVA and followed by Bonferroni post hoc tests. Data c was analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

Article Snippet: As shown, proBDNF protein did not affect T cell subgroups in normal mice (Fig. a–c).

Techniques: Mouse Assay, Injection, Flow Cytometry

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Systemic blockade of proBDNF restored meningeal pro-inflammatory microenvironment in septic mice. Mice were i.p. injected with proBDNF 30 min before LPS (5 mg kg −1 ) injection. Meninges were harvested 5 days after LPS injection for qPCR. a The level of CD4 gene expression was higher in the meninges of the McAb-proB group than in IgG controls in septic mice. b–f Gene levels were significantly lower in b IL-1β and c IL-6 but higher in d IL-4, e IFN-γ, and f IL-13 in the meninges after LPS injection in the McAb-proB group as compared to IgG control. n = 5 in each group. All experiments were performed at least in triplicate. Data were analyzed by unpaired T test, * P

Article Snippet: As shown, proBDNF protein did not affect T cell subgroups in normal mice (Fig. a–c).

Techniques: Mouse Assay, Injection, Real-time Polymerase Chain Reaction, Expressing

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Activation Regulates the Timing of the Maturation of Cortical Parvalbumin Interneuron Connectivity and Promotes Juvenile-like Plasticity in Adult Visual Cortex

doi: 10.1523/JNEUROSCI.2881-18.2019

Figure Lengend Snippet: proBNDF-mediated p75NTR activation in cortical PV cells reduces their perisomatic boutons. A , Experimental approach. B , The intensity of perisomatic PV immunostaining (green) is reduced in the binocular visual cortex ipsilateral to the minipump-releasing mut-proBDNF (Ipsi) compared with the contralateral cortex (Contra) in the same animal. On the other hand, perisomatic PV intensity in the ipsilateral cortex of PV_Cre;p75 flx/flx mice is similar to that observed in the contralateral, untreated cortex. C , Low ( C1 ) and high ( C2 ) magnification of PNN (red, WFA staining) enwrapping PV cells (green) shows a dramatic reduction in both PNN density and intensity in the visual cortex infused with mut-proBFNF. This effect is abolished in PV_Cre;p75 flx/flx mice. Scale bars: C1 , 100 μm; B , C2 , 10 μm. D , Quantification of the mean intensity of perisomatic PV-positive puncta in ipsilateral compared with contralateral cortex. I/C ratio is obtained for each animal and then averaged between different animals. Mean I/C ratio is significantly reduced in Mut-proBDNF-infused p75 Ctrl mice compared with Mut-proBDNF-infused PV_Cre;p75 flx/flx mice (unpaired t test, df = 8, t = 6.077, p = 0.0003). E , The ratio of mean PNN intensity around PV cells in ipsilateral versus contralateral cortex is significantly lower in p75 Ctrl than PV_Cre;p75 flx/flx mice infused with mut-proBDNF (unpaired t test, df = 8, t = 15.33, p

Article Snippet: Recombinant mouse proneurotrophin, proBDNF (wt-proBDNF, 10 ng/ml, Alomone Labs, catalog #B-240) and cleavage-resistant, recombinant mouse proBDNF (mut-proBDNF, 10 ng/ml, Alomone Labs, catalog #B-243) were, respectively, added with the culture medium during the specific time window indicated in Results.

Techniques: Activation Assay, Immunostaining, Mouse Assay, Staining

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Activation Regulates the Timing of the Maturation of Cortical Parvalbumin Interneuron Connectivity and Promotes Juvenile-like Plasticity in Adult Visual Cortex

doi: 10.1523/JNEUROSCI.2881-18.2019

Figure Lengend Snippet: Modulation of tPA activity affects the formation of PV cell innervations during early postnatal development. A , Control EP18 PV cell ( A1 , green represents Ctrl). B , PV cell treated with the tPA inhibitor PPACK from EP10–EP18 shows simpler axonal arborization, contacting less potential targets ( B2 , blue represents NeuN-positive somata). C , PV cell treated with tPA in the same time window shows a very complex axonal arbor ( C2 ) and an increase in both terminal branching and perisomatic boutons ( C3 , arrowheads) compared with control cells ( A2 , A3 ). D , PV cell treated simultaneously with tPA and mut-proBDNF shows axonal branching and perisomatic innervation more similar to those formed by PV cell treated with mut-proBDNF alone, suggesting that the effects of tPA application may be mediated by a decrease in endogenous proBDNF/mBDNF ratio. Stars indicate NeuN-positive somata that are not innervated. Scale bars: A1–D1 , 50 μm; A2–D2 , 10 μm; A3–D3 , 5 μm. E , Perisomatic boutons density (one-way ANOVA, F (3,20) = 121.2, p

Article Snippet: Recombinant mouse proneurotrophin, proBDNF (wt-proBDNF, 10 ng/ml, Alomone Labs, catalog #B-240) and cleavage-resistant, recombinant mouse proBDNF (mut-proBDNF, 10 ng/ml, Alomone Labs, catalog #B-243) were, respectively, added with the culture medium during the specific time window indicated in Results.

Techniques: Activity Assay

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Activation Regulates the Timing of the Maturation of Cortical Parvalbumin Interneuron Connectivity and Promotes Juvenile-like Plasticity in Adult Visual Cortex

doi: 10.1523/JNEUROSCI.2881-18.2019

Figure Lengend Snippet: proBNDF-mediated p75NTR activation in cortical PV cells restores ocular dominance plasticity in adult visual cortex in vivo . A , Typical VEP responses to the stimulation of either contralateral (blue) or ipsilateral (red) eye to the cortex in which the recording is performed in p75NTR Ctrl mice infused with either vehicle or mut-proBDNF, and PV_Cre;p75NTR flx/flx mice infused with mut-proBDNF. Calibration bars: 50 μV, 100 ms. B , C/I VEP ratio mean values. Three days of monocular deprivation do not affect the C/I VEP ratio in adult mice, although it leads to a significant decrease in the C/I VEP ratio in animals treated with mut-proBDNF. Mut-proBDNF effects are, however, abolished in PV_Cre;p75 flx/flx mice (one-way ANOVA, F (2,18) = 8.903, p = 0.0021). p75NTR Ctrl + vehicle: n = 9 mice; p75NTR Ctrl + mut-proBDNF: n = 5 mice; PV_Cre;p75 flx/flx +mut-proBDNF: n = 7 mice. C , ODI of p75NTR Ctrl mice infused with vehicle solution and PV_Cre;p75 flx/flx mice infused with mut-proBDNF are not significantly different from those of undeprived animals, whereas ODIs in p75 Ctrl mice treated with mut-proBDNF are significantly shifted toward the open eye (one-way ANOVA, F (2,443) = 5.203, p = 0.0058). D , Mean spontaneous discharge is significantly increased only in p75 Ctrl mice treated with mut-proBDNF (one-way ANOVA, F (2,443) = 4.580, p = 0.0107). p75NTR Ctrl + vehicle: n = 9 mice, 174 cells; p75NTR Ctrl + mut-proBDNF: n = 5 mice, 147 cells; PV_Cre;p75 flx/flx +mut-proBDNF: n = 6 mice, 125 cells. Gray area represents the C/I VEP ratio ( B ) or the ODI range ( C ) (mean ± SEM) in adult nondeprived animals ( n = 5 mice, 99 cells). * indicate p

Article Snippet: Recombinant mouse proneurotrophin, proBDNF (wt-proBDNF, 10 ng/ml, Alomone Labs, catalog #B-240) and cleavage-resistant, recombinant mouse proBDNF (mut-proBDNF, 10 ng/ml, Alomone Labs, catalog #B-243) were, respectively, added with the culture medium during the specific time window indicated in Results.

Techniques: Activation Assay, In Vivo, Mouse Assay

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Activation Regulates the Timing of the Maturation of Cortical Parvalbumin Interneuron Connectivity and Promotes Juvenile-like Plasticity in Adult Visual Cortex

doi: 10.1523/JNEUROSCI.2881-18.2019

Figure Lengend Snippet: mut-proBDNF destabilizes PV cell innervation, even after it has reached maturity. A , Control PV cell ( A1 , Ctrl, green) at EP32 with exuberant innervation field characterized by extensive branching contacting the majority of potential targets, dense boutons along axons ( A2 ), and terminal branches with prominent and clustered boutons ( A3 ; arrowheads) around NeuN-positive somata (blue). B , PV cell treated with wt-proBDNF from EP26-EP32 shows overall similar axon size ( B1 ), percentage of potentially targeted neurons ( B2 ), and perisomatic innervations ( B3 ) as control, untreated PV cells. C , PV cell treated with mut-proBDNF from EP26-EP32 shows a drastic reduction both in percentage of innervated cells ( C2 ) and perisomatic innervation ( C3 ). Stars indicate NeuN-positive somata that are not innervated. Scale bars: A1–C1 , 50 μm; A2–C2 , 10 μm; A3–C3 , 5 μm. D , Perisomatic bouton density (one-way ANOVA, F (2,18) = 93.34, p

Article Snippet: Recombinant mouse proneurotrophin, proBDNF (wt-proBDNF, 10 ng/ml, Alomone Labs, catalog #B-240) and cleavage-resistant, recombinant mouse proBDNF (mut-proBDNF, 10 ng/ml, Alomone Labs, catalog #B-243) were, respectively, added with the culture medium during the specific time window indicated in Results.

Techniques:

Journal: bioRxiv

Article Title: p75 Neurotrophin Receptor Regulates the Timing of the Maturation of Cortical Parvalbumin Cell Connectivity and Promotes Ocular Dominance Plasticity in Adult Visual Cortex

doi: 10.1101/392159

Figure Lengend Snippet: proBNDF-mediated p75NTR activation in cortical PV cells reduces their perisomatic boutons and restores ocular dominance plasticity in adult visual cortex in vivo . (a) Experimental approach. (b) The number of immunolabeled PV-positive puncta (green) surrounding NeuN-positive neuronal somata (red) is reduced in the binocular visual cortex ipsilateral to the minipump releasing mut-proBDNF (Ipsi) compared to the contralateral cortex (Contra) in the same animal. On the other hand, the number of PV-positive puncta per NeuN-positive profile in the ipsilateral cortex of PV-CRE; p75 flx/flx mice is similar to that observed in the contralateral, untreated cortex. (c) Low (c1) and high (c2) magnification of PNN (red, WFA staining) enwrapping PV cells (green) show a dramatic reduction in both PNN density and intensity in the visual cortex infused with mut-proBFNF. This effects is abolished in PV-CRE; p75 flx/flx mice. Scale bar, c1: 100µm; b, c2: 10µm. (d) Quantification of the mean number of PV-positive puncta per NeuN-positive profile in ipsilateral compared to contralateral cortex. Ipsi/Contra ratio is obtained for each animal, and then averaged between different animals. Mean Ipsi/Contra ratio is significantly reduced in Mut-proBDNF infused p75 Ctrl but not in PV-CRE; p75 flx/flx mice (t-test, p

Article Snippet: To label control PV cells, slices were transfected with PG67 _GFP bullets, while for the p75NTR-/- PV cells were transfected with both PG67 _GFP and PG67_Cre. wt-proBDNF and mut-proBDNF (10 ng/ml, Alomone Labs) were respectively added with the culture medium during the specific time window indicated in the results section.

Techniques: Activation Assay, In Vivo, Immunolabeling, Mouse Assay, Staining, T-Test

Journal: bioRxiv

Article Title: p75 Neurotrophin Receptor Regulates the Timing of the Maturation of Cortical Parvalbumin Cell Connectivity and Promotes Ocular Dominance Plasticity in Adult Visual Cortex

doi: 10.1101/392159

Figure Lengend Snippet: mut-proBDNF can destabilize PV cell innervation even after it has reached maturity. (a) Control PV cell (a1, Ctrl, green) at EP32 with exuberant innervation field characterized by extensive branching contacting the majority of potential targets, dense boutons along axons (a2), and terminal branches with prominent and clustered boutons (a3; arrowheads) around pyramidal cell somata (NeuN immunostaining, blue). (b) PV cell treated with wt-proBDNF from EP26-32 shows overall similar axon size (b1), percentage of potentially targeted neurons (B2) and perisomatic innervations (b3) as control, untreated PV cells. (c) PV cell treated with mut-proBDNF from EP26-32 shows a drastic reduction both in percentage of innervated cells (c2) and perisomatic innervation (c3). Stars indicate pyramidal cells somata that are not innervated. Scale bar, a1-c1: 50µm; a2-c2: 10µm; a3-c3: 5µm. (d) Perisomatic bouton density (e) terminal branching and (f) percentage of innervated cells of the three experimental groups. One-way Anova, post hoc Tukey test, p

Article Snippet: To label control PV cells, slices were transfected with PG67 _GFP bullets, while for the p75NTR-/- PV cells were transfected with both PG67 _GFP and PG67_Cre. wt-proBDNF and mut-proBDNF (10 ng/ml, Alomone Labs) were respectively added with the culture medium during the specific time window indicated in the results section.

Techniques: Immunostaining