Journal: ASN neuro

Article Title: Comparison of cortical and white matter traumatic brain injury models reveals differential effects in the subventricular zone and divergent Sonic hedgehog signaling pathways in neuroblasts and oligodendrocyte progenitors.

doi: 10.1177/1759091414551782

Figure Lengend Snippet: Figure 1. Shh-responsive cells heritably labeled after cortical injury from mild CCI. (a–e) Gli1-CreERT2;R26-IAP mice show strong AP labeling of multiple cell types in coronal brain sections. Mice were administered tamoxifen on Days 2 and 3 followed by survival until 2 weeks post-CCI (a, arrows indicate site of impact on dura) or postsham surgery (e). This heritable labeling identifies cells that were responding to high levels of Shh to drive Gli1 transcription during tamoxifen-induced nuclear translocation of the Cre-ER fusion protein. AP-labeled cells are abundant in the SVZ (arrowheads in b, c). Rare AP-labeled myelinating oligodendrocytes have processes that make characteristic T-intersections to extend along axons (arrows in b, c). AP-labeled cells with the morphology of astrocytes are common in the cerebral cortex (d). AP labeling is also observed in Shh-responsive endothelial cells of blood vessels (asterisks in c, d). Examples in (c) and (d) are from sections adjacent to (a) and (b). (f–h) Gli1-CreERT2;R26-YFP mice show a similar pattern of heritable YFP labeling as with AP at 2 weeks postsurgery in sham (e, f) and CCI mice (a, g). GFAP immunolabeling demonstrates an astrocytic reaction to craniotomy in sham mice (g). GFAP does not consistently label the population of cortical astrocytes that express YFP (g). Cortical astrocytes with thin, highly branched processes are double labeled with YFP and S100b (g, inset). Not all S100b cells are labeled with YFP, particularly reactive astrocytes in the lesion and penumbra (g, arrows indicate site of impact on dura). YFP cells that remain in the lesion and penumbra maintain a morphology with fine, complex processes that is distinct from the reactive astrocytes labeled for S100b but not YFP (h). Nuclear DAPI colocalization was used to differentiate intact YFP cells from debris or autofluorescence in lesions (h). (i) At both 2 and 6 weeks post-CCI, YFP-labeled cells are reduced in the cortex under the site of impact (injury effect, **p ¼.0023, n ¼ 4 mice per condition). cc ¼ corpus callosum, ac ¼ anterior commissure. Scale bars: a ¼ 1 mm; b, e ¼ 500 mm; c, d ¼ 50 mm; f to h ¼ 250 mm.

Article Snippet: Secondary antibodies used were as follows: donkey antirabbit IgG F(ab0)2 conjugated with Cy3 (Jackson ImmunoResearch, West Grove, PA) to detect bAPP; goat anti-rabbit IgG conjugated with Alexa Fluor 488 (Life Technologies, Grand Island, NY) to detect DCX, NG2, GFAP, and Olig2; donkey anti-mouse IgG F(ab0)2 conjugated with Cy3 (Jackson ImmunoResearch) to detect S100b; donkey anti-sheep IgG conjugated with Cy3 (Jackson ImmunoResearch) to detect EGFR; and goat anti-rat IgG conjugated with Alexa Fluor 555 (Life Technologies) to detect YFP.

Techniques: Labeling, Translocation Assay, Immunolabeling

Journal: Neuron

Article Title: Driving axon regeneration by orchestrating neuronal and non-neuronal innate immune responses via the IFNγ-cGAS-STING axis

doi: 10.1016/j.neuron.2022.10.028

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti-S100β , Cell Signaling , Cat#9550; RRID: AB_10949319.

Techniques: Control, Virus, Plasmid Preparation, Recombinant, RNAscope, Multiplex Assay, Fluorescence, Reverse Transcription, SYBR Green Assay, Single Cell Gel Electrophoresis, Knock-Out, Knock-In, shRNA, Sequencing, Software

Journal: International Journal of Molecular Sciences

Article Title: S100B Protein Stimulates Proliferation and Angiogenic Mediators Release through RAGE/pAkt/mTOR Pathway in Human Colon Adenocarcinoma Caco-2 Cells

doi: 10.3390/ijms20133240

Figure Lengend Snippet: S100B stimulates Caco-2 cell proliferation, migration and invasion and its effect is blocked by S100B monoclonal antibody (mAb). ( A ) 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay showing the effect of S100B (0.005–5 µM) in the presence or absence of S100BmAb (1:10 5 –1.10 4 v / v diluted), receptor for advanced glycation end products (RAGE)mAB (1:10 4 v / v diluted) and/or SB203580 p38/pAkt inhibitor (10 μM) on Caco-2 cell proliferation rate at 48 h. ( B ) Wound healing assay and ( D ) the relative quantification indicating a concentration-dependent inhibitory effect of S100BmAb on cellular migration induced by S100B (0.05–5 µM). The graphs show also RAGEmAb (1:10 4 v / v diluted) and/or SB203580 p38/pAkt inhibitor (10 μM) against S100B 5 μM stimulus at 48 h. ( C ) Cell invasion assay and ( E ) the relative quantification of invading cells following S100B (0.05–5 µM) exposure and relative inhibitory effect of S100BmAb (1:10 5 –1:10 4 v / v diluted). Figures also show the effect of RAGEmAb (1:10 4 v / v diluted) and SB203580 p38/pAkt inhibitor (10 μM) versus S100B 5 μM stimulus. Results were expressed as mean ± standard error (SEM) of n = 6 experiments performed in triplicate. * p < 0.05; ** p < 0.01 and *** p < 0.001 versus vehicle; °° p < 0.01 and °°° p < 0.001 versus S100B 5 μM; # p < 0.05 and § p < 0.05 respectively versus S100B 5 μM-treated cells. Scale bar: 100 µm; Magnification 10X.

Article Snippet: S100B protein was from Sigma-Aldrich (St. Louis, MO, USA) and mouse monoclonal S100B antibody (S100BmAb) was purchased from AbCam (Cambridge, UK).

Techniques: Migration, MTT Assay, Wound Healing Assay, Concentration Assay, Invasion Assay

Journal: International Journal of Molecular Sciences

Article Title: S100B Protein Stimulates Proliferation and Angiogenic Mediators Release through RAGE/pAkt/mTOR Pathway in Human Colon Adenocarcinoma Caco-2 Cells

doi: 10.3390/ijms20133240

Figure Lengend Snippet: S100B stimulates pro-angiogenic vascular endothelial growth factor (VEGF) and NO release through Akt/mammalian target of rapamycin (mTOR) pathways and relative molecular downstream regulation by S100BmAb. ( A ) Immunoreactive bands referred to RAGE, phosphorylated/unphosphorylated-p38MAPK, phosphorylated/unphosphorylated Akt, phosphorylated ERK/unphosphorylated ERK, phosphorylated/unphosphorylated-mTOR, wild type (wt)p53, hypoxia-inducible factor 1-alpha (HIF1α) protein expression and ( H ) their relative densitometric analysis (arbitrary units normalized on the expression of the housekeeping protein β-actin, showing the effect of S100B (0.05–5 µM) in the presence of S100BmAb (1:10 5 –1:10 4 v / v diluted) at 24 h, S100BmAb alone (1:10 4 v / v diluted) or S100B 5 μM in the presence of, respectively RAGEmAb (1:10 4 v / v diluted) or SB203580 (10 μM). ( B ) Immunofluorescence analysis showing inducible nitric oxide-synthase (iNOS; green), VEGF-R1 and ( C ) VEGF-R2 (red) and ( D ) Ki67 (red) immunoreactivity with ( E ) the relative quantification of iNOS (green bars), VEGF-R1 (yellow bars) VEGF-R2 (red bars) and Ki67 (blue bars) protein expression in Caco-2 cells treated with S100B (5 µM) in presence or absence of S100BmAb (1:10 4 v / v diluted), or RAGEmAb (1:10 4 v / v diluted) and SB203580 (10 μM). In the same experimental conditions, ( F and G ) quantification of nitrite and VEGF levels respectively in the supernatant media of Caco-2 cells exposed to the same treatment. Results were expressed as mean ± SEM of n = 6 experiments performed in triplicate.* p < 0.05; ** p < 0.01 and *** p < 0.001 versus vehicle; ° p < 0.05, °° p < 0.01 and °°° p < 0.001 versus S100B 5 μM; # p < 0.05 ## p < 0.01; § p < 0.05 and §§ p < 0.01, respectively versus S100B 5 μM-treated cells. Scale bar: 10 µm; Magnification 10X.

Article Snippet: S100B protein was from Sigma-Aldrich (St. Louis, MO, USA) and mouse monoclonal S100B antibody (S100BmAb) was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: S100B Protein Stimulates Proliferation and Angiogenic Mediators Release through RAGE/pAkt/mTOR Pathway in Human Colon Adenocarcinoma Caco-2 Cells

doi: 10.3390/ijms20133240

Figure Lengend Snippet: Representation of S100B effects on Caco-2 cells. ( A ) S100B interacts at RAGE receptors and downstream activates p38/Akt signaling pathway leading to cell proliferation, increased release of cell migration and angiogenesis. ( B ) S100B effects are blocked by specific S100BmAb, as well as, RAGE specific antibody and p38/Akt inhibitor SB203580, leading to a significant control of angiogenesis and cell invasion in vitro.

Article Snippet: S100B protein was from Sigma-Aldrich (St. Louis, MO, USA) and mouse monoclonal S100B antibody (S100BmAb) was purchased from AbCam (Cambridge, UK).

Techniques: Migration, In Vitro

Journal: Molecular Autism

Article Title: Neuroglia in the autistic brain: evidence from a preclinical model

doi: 10.1186/s13229-018-0254-0

Figure Lengend Snippet: Primer sequences and general conditions used to perform real-time qPCR

Article Snippet: Rabbit α-S100B , Genetex , 1:1000 , HRP conjugated goat anti-rabbit IgG 1:10000 , Jackson ImmunoResearch.

Techniques:

Journal: Molecular Autism

Article Title: Neuroglia in the autistic brain: evidence from a preclinical model

doi: 10.1186/s13229-018-0254-0

Figure Lengend Snippet: Experimental conditions used to perform western blot experiments

Article Snippet: Rabbit α-S100B , Genetex , 1:1000 , HRP conjugated goat anti-rabbit IgG 1:10000 , Jackson ImmunoResearch.

Techniques: Western Blot

Journal: Molecular Autism

Article Title: Neuroglia in the autistic brain: evidence from a preclinical model

doi: 10.1186/s13229-018-0254-0

Figure Lengend Snippet: Effect of prenatal VPA exposure on astrocytes in infancy. Analysis of the neurotrophin S100B and the GFAP in the PfC, Cb, and HPC of healthy- (open bars, Veh) and autistic-like (black bars, VPA) infant rats (PND 13). Representation of the relative concentration of S100B ( a ) and GFAP ( c ) in VPA animals compared to control (Veh), normalized to both TBP and HPRT (ΔΔCq; N = 3, in triplicate). Representative western blots for S100B ( b ) and GFAP ( d ) proteins, and densitometric analyses are normalized to β-actin used as loading control. Results are expressed as percentage of control (Veh) ( N = 3, in triplicate). Representative fluorescence micrographs of GFAP (green) staining in the PfCGL and MLof Cb, and CA1, CA2, CA3, and the hilus of the DG of the HPC. Nuclei were stained with Hoechst (blue) ( e ). The images have been analyzed by counting the number of GFAP-positive cells in 2.4 × 10 5 μm 3 (scale bar 50 μm; N = 3, 4 times) ( f ). All data are presented as means ± SEM. Statistical analysis was performed by t test (* p < 0.05; *** p < 0.001 vs Veh group)

Article Snippet: Rabbit α-S100B , Genetex , 1:1000 , HRP conjugated goat anti-rabbit IgG 1:10000 , Jackson ImmunoResearch.

Techniques: Concentration Assay, Control, Western Blot, Fluorescence, Staining

Journal: Molecular Autism

Article Title: Neuroglia in the autistic brain: evidence from a preclinical model

doi: 10.1186/s13229-018-0254-0

Figure Lengend Snippet: Effect of the prenatal VPA exposure on astrocytes in adolescence. Evaluation of the neurotrophin S100B and the cytoskeletal GFAP in the PfC, Cb, and HPC of healthy- (open bars, Veh) and autistic-like (black bars, VPA) adolescent rats (PND 35). Representation of the relative concentration of S100B ( a ) and GFAP ( c ) in VPA animals compared to control (Veh), normalized to both TBP and HPRT (ΔΔCq; N = 3, in triplicate). Representative western blots for S100B ( b ) and GFAP ( d ) proteins, and densitometric analyses normalized to β-actin used as loading control. Results are expressed as percentage of control (Veh) ( N = 3, in triplicate). Representative fluorescence micrographs of GFAP (green) staining in the PfC, GL and ML of Cb, and stratum radiatum of CA1, CA2, CA3, and the hilus of the DG of the HPC. Nuclei were stained with Hoechst (blue) ( e ). The images have been analyzed by counting the number of GFAP-positive cells in 2.4 × 10 5 μm 3 (scale bar 50 μm; N = 3, 4 times) ( f ). All data are presented as means ± SEM. Statistical analysis was performed by t test (* p < 0.05; ** p < 0.01; *** p < 0.001 vs Veh group)

Article Snippet: Rabbit α-S100B , Genetex , 1:1000 , HRP conjugated goat anti-rabbit IgG 1:10000 , Jackson ImmunoResearch.

Techniques: Concentration Assay, Control, Western Blot, Fluorescence, Staining

Journal: Molecular Autism

Article Title: Neuroglia in the autistic brain: evidence from a preclinical model

doi: 10.1186/s13229-018-0254-0

Figure Lengend Snippet: Effect of the prenatal VPA exposure on astrocytes in adulthood. Evaluation of the neurotrophin S100B and the cytoskeletal GFAP in the PfC, Cb, and HPC of healthy- (open bars, Veh) and autistic-like (black bars, VPA) adult rats (PND 90). Representation of the relative concentration of S100B ( a ) and GFAP ( c ) in VPA animals compared to control (Veh), normalized to both TBP and HPRT (ΔΔCq; N = 3, in triplicate). Representative western blots for S100B ( b ) and GFAP ( d ) proteins, and densitometric analyses normalized to β-actin used as loading control. Results are expressed as percentage of control (Veh) ( N = 3, in triplicate). Representative fluorescence micrographs of GFAP (green) staining in the PfC, GL and ML of Cb, and stratum radiatum of CA1, CA2, CA3, and the hilus of the DG of the HPC. Nuclei were stained with Hoechst (blue) ( e ). The images have been analyzed by counting the number of GFAP-positive cells in 2.4 × 10 5 μm 3 (scale bar 50 μm; N = 3, 4 times) ( f ). All data are presented as means ± SEM. Statistical analysis was performed by t test (* p < 0.05; *** p < 0.001 vs Veh group)

Article Snippet: Rabbit α-S100B , Genetex , 1:1000 , HRP conjugated goat anti-rabbit IgG 1:10000 , Jackson ImmunoResearch.

Techniques: Concentration Assay, Control, Western Blot, Fluorescence, Staining