neurotrophic factor Search Results


95
Alomone Labs neurotrophic factor gdnf
Neurotrophic Factor Gdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Neurotech Pharmaceuticals Inc ciliary neurotrophic factor
Ciliary Neurotrophic Factor, supplied by Neurotech Pharmaceuticals Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Elabscience Biotechnology elisa kit
Fig. 2 Riluzole treatment restores <t>hippocampal</t> <t>BDNF</t> in the irradiated hippocampus. 10–12 weeks old WT male mice received cranial radiation therapy (RT) followed by riluzole (RZ) treatment (13 mg/kg) in drinking water for 6–7 weeks. An <t>ELISA-based</t> quantification of BDNF from the micro-dissected hippocampus showed RT-induced reductions in the RT + Vehicle group. Importantly, RZ treatment in the cranially irradiated mice showed significant restoration of BDNF levels. Data are presented as mean ± SEM (N = 6–10 mice per group). P values were derived from two-way ANOVA and Bonferroni’s multiple comparisons test
Elisa Kit, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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93
Alomone Labs human mbdnf
( A ) Representative Western blot (a) and their semi-quantitative analyses of mature BDNF <t>(b),</t> <t>proBDNF</t> (c) and their ratio (d) in the local tissue after 10 μL 5% formalin intra-plantar injection into Kunming mice (*p < 0.05, **p < 0.01 versus control, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). ( B ) H&E staining (a and d) and immunohistochemsitry (b,c and e–g) of proBDNF in the foot skin at 3 h post-formalin injection. proBDNF is expressed in the epidermis, basal layer and subcutaneous layers in the foot skin (b,c); Higher magnification (box in b) showing proBDNF is also mildly expressed in the nerve fibers in the control plantar (c); Responding to peripheral inflammation by 5% formalin intra-plantar injection, intensive proBDNF immunoreactivity is observed and mainly localized in the inflammatory cells (f, black arrows) and nerve fiber-like structures (g). Scale bars: 50 μm, 3 replicates, n = 3 per group. ( C ) a, Representative Western blot of proBDNF and <t>mBDNF;</t> b–d, Semi-quantitative analyses of mBDNF, proBDNF and their ratio in the inflamed tissue after Complete Freund Adjuvant (CFA, 10 μL) intra-plantar injection into Kunming mice (*p < 0.05, **p < 0.01versus control, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). ( D ) Histological staining (a) and proBDNF immunohistochemistry (b,c) in the plantar at 1 day post-CFA injection; c, higher magnification of box in b showing that proBDNF is highly expressed in the inflammatory cells. Scale bar, 100 μm, 3 replicates, n = 3 per group. Data bars represent mean ± s.e.m.
Human Mbdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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95
Alomone Labs rabbit anti bdnf
(A) Representative images <t>showing</t> <t>GFP</t> and c-Fos (arrows) expression patterns in the PVT, a downstream target of the VMH, in AAV-DIO-EYFP, AAV-DIO-NaChBac+sgRNA, AAV-DIO-NaChBac+sgVglut2 injection SF1-Cre::ERa-Cre mice. scale bar=250 um. (B) Representative images showing GFP (green) and c-Fos (red, arrows) expression patterns in the PVT, a downstream target of the VMH, in SF1-Cre::ERa-Cre::BDNFflox/++EYFP, SF1-Cre::ERa-Cre::BDNFflox/++NaChBac, SF1-Cre::ERa-Cre::BDNFflox/flox+NaChBac mice. scale bar=250 um. (C) Number of c-Fos positive cells in the PVT region related to panel A. n=4-5 per group; one-way ANOVA; ***P<0.001 versus EYFP; ###P<0.001 versus NaChBac+sgRNA. (D) number of c-Fos positive cells in the PVT region related to panel A. n=4-5 per group; one-way ANOVA; ***P<0.001 versus EYFP; ###P<0.001 versus SF1-Cre::ERa-Cre:: BDNFflox/flox+NaChBac. (E) Diagram of viral delivery (AAV-DIO-mCherry, <t>AAV-DIO-BDNF-mCherry+sgRNA,</t> AAV-DIO-BDNF-mCherry+sgVglut2) into the VMH of SF1-Cre::ERa-Cre mice. (F) Body weight trajectories of SF1-Cre::ERa-Cre mice after VH injection of virus. n=4 (EYFP, BDNF+sgVglut2), n=5 (BDNF+sgRNA); two-way ANOVA; *P<0.05, **P<0.01, ***P<0.001 versus mCherry; #P<0.05 versus BDNF+sgRNA. (G) Daily food intake. n=4-5 per group, one-way ANOVA, **P=0.006 versus mCherry; #P=0.043 versus BDNF+sgRNA. (H) Total number of jumps during a 10-min test. n=4-5 per group; one-way ANOVA; ***P<0.01 versus mCherry; ###P<0.001 versus BDNF+sgRNA. (I, J) time spent in the light room in light-dark test (I), social time with stranger in three-chamber test (J). n=4-5 per group; one-way ANOVA; *P<0.05, ***P<0.01 versus mCherry; #P<0.05 versus BDNF+sgRNA. (K) Representative images showing mCherry (green) and c-Fos (magenta, arrows) expression patterns in the PVT, a downstream of VMH, in mCherry, BDNF+sgRNA, BDNF+sgVglut2 injection SF1-Cre::ERa-Cre mice. scale bar=250 um. (L) number of c-Fos positive cells in PVT region related to panel K. n=4-5 per group; one-way ANOVA; ***P<0.001 versus mCherry; ###P<0.001 versus BDNF+sgRNA.
Rabbit Anti Bdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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94
Alomone Labs cholesterol
FIGURE 1 (a) Workflow of data mining. The TMR (transmembrane sequence +5 amino acid residues from each N- and C-terminal sides) of curated entries found in UniProt using the code for tyrosine-kinase receptor family (2.7.10.1) were screened for the presence of <t>cholesterol</t> recognition and alignment consensus (CRAC and CARC). The incidence of CRAC and CARC motifs in (b) human, (c) mouse, (d) zebrafish, (e) fruit fly, and (f) C. elegans TMR of RTK family members. (g) Library of CRAC and CARC sequences (all combinations used can be found in the stored data). Residue sequence predicted to be embedded into the cell membrane shown in yellow
Cholesterol, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/neurotrophic+factor/human+BDNF-Biotin/pm33825223-77-30-24
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91
Alomone Labs anti trkb
FIGURE 1 (a) Workflow of data mining. The TMR (transmembrane sequence +5 amino acid residues from each N- and C-terminal sides) of curated entries found in UniProt using the code for tyrosine-kinase receptor family (2.7.10.1) were screened for the presence of <t>cholesterol</t> recognition and alignment consensus (CRAC and CARC). The incidence of CRAC and CARC motifs in (b) human, (c) mouse, (d) zebrafish, (e) fruit fly, and (f) C. elegans TMR of RTK family members. (g) Library of CRAC and CARC sequences (all combinations used can be found in the stored data). Residue sequence predicted to be embedded into the cell membrane shown in yellow
Anti Trkb, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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93
Alomone Labs recombinant human bdnf protein rhbdnf
Western blot analysis of the <t>BDNF</t> ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the prefrontal (prelimbic/infralimbic) cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. ***: p < 0.001 (post hoc Duncan’s multiple range test).
Recombinant Human Bdnf Protein Rhbdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/neurotrophic+factor/Recombinant+human+BDNF+protein/pmc12384143-180-17-24
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Cell Signaling Technology Inc snai1
Fig. 3. Suppression of treRNA by miR-190a inhibits EMT in HCC. (A) 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or sitreRNA2), 20 nM miR-190a, or NC RNA was transfected into Huh-7 or HepG2 cells. EMT related genes were quantified 72 h post-transfection as indicated. (B) Cells were treated with antagomiR (anti-miR- 190a) or NC RNA as in (A). The mRNA level of different EMT genes was quantified and normalized to b-actin. (C) Cell lysates were blotting with EMT marker E-cadherin (E-cad), Claudin-1 (Claud), Vimentin (Vimen), N-cadherin (N-cad), <t>SNAI1</t> (Snail). b-Actin was used as endogenous control. The EMT marker expression level in was calculated using Image J and presented as histogram (right panel). (D) Cells were treated with TGF-b for 12 h and transfected with 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or si-treRNA2) or NC RNA subsequently. The morphological change was captured 24 h after transfection.
Snai1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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94
Cusabio immunoassay kit
Fig. 3. Suppression of treRNA by miR-190a inhibits EMT in HCC. (A) 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or sitreRNA2), 20 nM miR-190a, or NC RNA was transfected into Huh-7 or HepG2 cells. EMT related genes were quantified 72 h post-transfection as indicated. (B) Cells were treated with antagomiR (anti-miR- 190a) or NC RNA as in (A). The mRNA level of different EMT genes was quantified and normalized to b-actin. (C) Cell lysates were blotting with EMT marker E-cadherin (E-cad), Claudin-1 (Claud), Vimentin (Vimen), N-cadherin (N-cad), <t>SNAI1</t> (Snail). b-Actin was used as endogenous control. The EMT marker expression level in was calculated using Image J and presented as histogram (right panel). (D) Cells were treated with TGF-b for 12 h and transfected with 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or si-treRNA2) or NC RNA subsequently. The morphological change was captured 24 h after transfection.
Immunoassay Kit, supplied by Cusabio, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/neurotrophic+factor/Rat+Ciliary+Neurotrophic+Factor%2CCNTF+ELISA+Kit/pm41856440-99-41-50
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Cusabio cntf
Fig. 3. Suppression of treRNA by miR-190a inhibits EMT in HCC. (A) 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or sitreRNA2), 20 nM miR-190a, or NC RNA was transfected into Huh-7 or HepG2 cells. EMT related genes were quantified 72 h post-transfection as indicated. (B) Cells were treated with antagomiR (anti-miR- 190a) or NC RNA as in (A). The mRNA level of different EMT genes was quantified and normalized to b-actin. (C) Cell lysates were blotting with EMT marker E-cadherin (E-cad), Claudin-1 (Claud), Vimentin (Vimen), N-cadherin (N-cad), <t>SNAI1</t> (Snail). b-Actin was used as endogenous control. The EMT marker expression level in was calculated using Image J and presented as histogram (right panel). (D) Cells were treated with TGF-b for 12 h and transfected with 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or si-treRNA2) or NC RNA subsequently. The morphological change was captured 24 h after transfection.
Cntf, supplied by Cusabio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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93
Cusabio elisa kit
Cellular localization and uptake of fluorescently labeled sEVs with SCs. (A) Cellular colocalization: SP8 was used to photograph sEVs prelabeled with PKH67 (green fluorescence) with Hoechst 33,342 (blue fluorescence)-stained SC nuclei. (B) Western blot: Detection of ERK1/2, ZEB2, and c-JUN expression levels in NC Schwann cells and after 48-h treatment with hypoxia sEVs. (C) Real-time PCR: Inflammatory and other restoration-related factors (IL-1β, IL-6, TNF-α, etc.) were detected. (D) <t>ELISA</t> <t>test:</t> <t>GDNF</t> NDF and NT-3 in the supernatant of Schwann cell preparation. Statistical significance, * p < 0.05,** p < 0.01, and **** p < 0.0001. *Significant difference.
Elisa Kit, supplied by Cusabio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Fig. 2 Riluzole treatment restores hippocampal BDNF in the irradiated hippocampus. 10–12 weeks old WT male mice received cranial radiation therapy (RT) followed by riluzole (RZ) treatment (13 mg/kg) in drinking water for 6–7 weeks. An ELISA-based quantification of BDNF from the micro-dissected hippocampus showed RT-induced reductions in the RT + Vehicle group. Importantly, RZ treatment in the cranially irradiated mice showed significant restoration of BDNF levels. Data are presented as mean ± SEM (N = 6–10 mice per group). P values were derived from two-way ANOVA and Bonferroni’s multiple comparisons test

Journal: Acta neuropathologica communications

Article Title: BDNF augmentation reverses cranial radiation therapy-induced cognitive decline and neurodegenerative consequences.

doi: 10.1186/s40478-024-01906-9

Figure Lengend Snippet: Fig. 2 Riluzole treatment restores hippocampal BDNF in the irradiated hippocampus. 10–12 weeks old WT male mice received cranial radiation therapy (RT) followed by riluzole (RZ) treatment (13 mg/kg) in drinking water for 6–7 weeks. An ELISA-based quantification of BDNF from the micro-dissected hippocampus showed RT-induced reductions in the RT + Vehicle group. Importantly, RZ treatment in the cranially irradiated mice showed significant restoration of BDNF levels. Data are presented as mean ± SEM (N = 6–10 mice per group). P values were derived from two-way ANOVA and Bonferroni’s multiple comparisons test

Article Snippet: BDNF levels were quantified using a commercially available ELISA kit (E-EL-M0203, Elabscience Biotechnology) and uncoated ELISA plates (Nunc MaxiSorp, Biolegend).

Techniques: Irradiation, Enzyme-linked Immunosorbent Assay, Derivative Assay

( A ) Representative Western blot (a) and their semi-quantitative analyses of mature BDNF (b), proBDNF (c) and their ratio (d) in the local tissue after 10 μL 5% formalin intra-plantar injection into Kunming mice (*p < 0.05, **p < 0.01 versus control, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). ( B ) H&E staining (a and d) and immunohistochemsitry (b,c and e–g) of proBDNF in the foot skin at 3 h post-formalin injection. proBDNF is expressed in the epidermis, basal layer and subcutaneous layers in the foot skin (b,c); Higher magnification (box in b) showing proBDNF is also mildly expressed in the nerve fibers in the control plantar (c); Responding to peripheral inflammation by 5% formalin intra-plantar injection, intensive proBDNF immunoreactivity is observed and mainly localized in the inflammatory cells (f, black arrows) and nerve fiber-like structures (g). Scale bars: 50 μm, 3 replicates, n = 3 per group. ( C ) a, Representative Western blot of proBDNF and mBDNF; b–d, Semi-quantitative analyses of mBDNF, proBDNF and their ratio in the inflamed tissue after Complete Freund Adjuvant (CFA, 10 μL) intra-plantar injection into Kunming mice (*p < 0.05, **p < 0.01versus control, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). ( D ) Histological staining (a) and proBDNF immunohistochemistry (b,c) in the plantar at 1 day post-CFA injection; c, higher magnification of box in b showing that proBDNF is highly expressed in the inflammatory cells. Scale bar, 100 μm, 3 replicates, n = 3 per group. Data bars represent mean ± s.e.m.

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: ( A ) Representative Western blot (a) and their semi-quantitative analyses of mature BDNF (b), proBDNF (c) and their ratio (d) in the local tissue after 10 μL 5% formalin intra-plantar injection into Kunming mice (*p < 0.05, **p < 0.01 versus control, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). ( B ) H&E staining (a and d) and immunohistochemsitry (b,c and e–g) of proBDNF in the foot skin at 3 h post-formalin injection. proBDNF is expressed in the epidermis, basal layer and subcutaneous layers in the foot skin (b,c); Higher magnification (box in b) showing proBDNF is also mildly expressed in the nerve fibers in the control plantar (c); Responding to peripheral inflammation by 5% formalin intra-plantar injection, intensive proBDNF immunoreactivity is observed and mainly localized in the inflammatory cells (f, black arrows) and nerve fiber-like structures (g). Scale bars: 50 μm, 3 replicates, n = 3 per group. ( C ) a, Representative Western blot of proBDNF and mBDNF; b–d, Semi-quantitative analyses of mBDNF, proBDNF and their ratio in the inflamed tissue after Complete Freund Adjuvant (CFA, 10 μL) intra-plantar injection into Kunming mice (*p < 0.05, **p < 0.01versus control, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). ( D ) Histological staining (a) and proBDNF immunohistochemistry (b,c) in the plantar at 1 day post-CFA injection; c, higher magnification of box in b showing that proBDNF is highly expressed in the inflammatory cells. Scale bar, 100 μm, 3 replicates, n = 3 per group. Data bars represent mean ± s.e.m.

Article Snippet: For ELISA assay, 96-well polystyrene microtiter plates (Nunc, Roskilde, Denmark, USA) were coated with the generated human proBDNF prodomain, and commercial human, rat and mice proBDNF proteins (Alomone Labs, Israel), and human mBDNF (Alomone Labs, Israel) (all at 1 μg/mL with total volume 50 μL) overnight at 4 °C.

Techniques: Western Blot, Injection, Control, Comparison, Staining, Adjuvant, Immunohistochemistry

( A ) Dosage effect of exogenous proBDNF protein on PWT by injection of proBDNF protein into the plantar (*P < 0.05, **p < 0.01 versus baseline, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 10–12 per group). ( B ) Ectopic overexpression of proBDNF by intra-plantar injection of Ad-proBDNF or Ad-EGFP reduces PWT dramatically in Kunming mice. (a) Representative proBDNF Western blot (upper panel) and its semi-quantitative analysis (lower panel, *P < 0.05, ***p < 0.001 versus control, ### p < 0.001 versus indicated groups, one-way ANOVA followed by Tukey’s Multiple Comparison post hoc test, n = 4 per group); (b) Representative fluorescent images after delivery of Ad-EGFP . Scale bar, 50 μm, 3 replicates, n = 3 per group; (c) PWT at 7 days post-injection of Ad-proBDNF or Ad-EGFP control (***p < 0.001 versus Ad-EGFP , student’s t test, n = 7 per group). ( C ) Co-injection of proBDNF (0.1 μg), but not mBDNF (0.1 μg) restored the biphasic nociceptive response after low-concentration of formalin (0.5%) intra-plantar injection (*p < 0.05versus vehicle, two-way ANOVA followed by Bonferroni’s Multiple Comparison post hoc test, n = 10–12 per group). ( D ) Exogenous proBDNF (1 μg) intra-plantar injection induces ERK activation in the ipsilateral spinal cord dorsal horn at 3 h post-injection, Scale bar, 50 μm, n = 3 per group, 3 replicates. ( E ) Spinal p-ERK expression at 3 h after proBDNF (1 μg) intra-plantar injection. (a) Representative Western blot and (b,c) their semi-quantitative analyses of p-ERK, (*p < 0.05, **p < 0.01 versus proBDNF-R, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). Data bars represent mean ± s.e.m.

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: ( A ) Dosage effect of exogenous proBDNF protein on PWT by injection of proBDNF protein into the plantar (*P < 0.05, **p < 0.01 versus baseline, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 10–12 per group). ( B ) Ectopic overexpression of proBDNF by intra-plantar injection of Ad-proBDNF or Ad-EGFP reduces PWT dramatically in Kunming mice. (a) Representative proBDNF Western blot (upper panel) and its semi-quantitative analysis (lower panel, *P < 0.05, ***p < 0.001 versus control, ### p < 0.001 versus indicated groups, one-way ANOVA followed by Tukey’s Multiple Comparison post hoc test, n = 4 per group); (b) Representative fluorescent images after delivery of Ad-EGFP . Scale bar, 50 μm, 3 replicates, n = 3 per group; (c) PWT at 7 days post-injection of Ad-proBDNF or Ad-EGFP control (***p < 0.001 versus Ad-EGFP , student’s t test, n = 7 per group). ( C ) Co-injection of proBDNF (0.1 μg), but not mBDNF (0.1 μg) restored the biphasic nociceptive response after low-concentration of formalin (0.5%) intra-plantar injection (*p < 0.05versus vehicle, two-way ANOVA followed by Bonferroni’s Multiple Comparison post hoc test, n = 10–12 per group). ( D ) Exogenous proBDNF (1 μg) intra-plantar injection induces ERK activation in the ipsilateral spinal cord dorsal horn at 3 h post-injection, Scale bar, 50 μm, n = 3 per group, 3 replicates. ( E ) Spinal p-ERK expression at 3 h after proBDNF (1 μg) intra-plantar injection. (a) Representative Western blot and (b,c) their semi-quantitative analyses of p-ERK, (*p < 0.05, **p < 0.01 versus proBDNF-R, one-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test, n = 3 per group). Data bars represent mean ± s.e.m.

Article Snippet: For ELISA assay, 96-well polystyrene microtiter plates (Nunc, Roskilde, Denmark, USA) were coated with the generated human proBDNF prodomain, and commercial human, rat and mice proBDNF proteins (Alomone Labs, Israel), and human mBDNF (Alomone Labs, Israel) (all at 1 μg/mL with total volume 50 μL) overnight at 4 °C.

Techniques: Injection, Comparison, Over Expression, Western Blot, Control, Concentration Assay, Activation Assay, Expressing

( A ) ELISA assay for the immunoreactivity of 2B11 against human proBDNF prodomain, and human, rat and mice proBDNF proteins, and human mature BDNF (mBDNF). 2B11 has strong immunoreactivity against proBDNF and prodomain, but not mBDNF; ( B ) Representative Western blot of human proBDNF and mBDNF detected by 2B11 (dilution 1:2000), note that 2B11 specifically recognizes proBDNF, but not mBDNF. ( C ) Representative images of neurosphere radiant migration treated by proBDNF, mBDNF, sheep polyclonal anti-proBDNF antibody, mouse monoclonal anti-proBDNF antibody 2B11 and co-treatment. ( D ) Statistical analysis of neurosphere migration radiance assay (***P < 0.001 versus control, # p < 0.05 versus indicated group, one-way ANOVA followed by Tukey’s Multiple Comparison post hoc test). Neurospheres treated with proBDNF (100 ng/mL) showed dormancy without any neuronal migration and neurospheres had no morphological changes. Neurospheres treated with 2B11 (100 ng/ml) showed strong migration capability comparing with other groups. Neurospheres treated with 2B11 and proBDNF (100 ng/ml) showed similar ability of migration with sheep anti-proBDNF antibody treatment group.

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: ( A ) ELISA assay for the immunoreactivity of 2B11 against human proBDNF prodomain, and human, rat and mice proBDNF proteins, and human mature BDNF (mBDNF). 2B11 has strong immunoreactivity against proBDNF and prodomain, but not mBDNF; ( B ) Representative Western blot of human proBDNF and mBDNF detected by 2B11 (dilution 1:2000), note that 2B11 specifically recognizes proBDNF, but not mBDNF. ( C ) Representative images of neurosphere radiant migration treated by proBDNF, mBDNF, sheep polyclonal anti-proBDNF antibody, mouse monoclonal anti-proBDNF antibody 2B11 and co-treatment. ( D ) Statistical analysis of neurosphere migration radiance assay (***P < 0.001 versus control, # p < 0.05 versus indicated group, one-way ANOVA followed by Tukey’s Multiple Comparison post hoc test). Neurospheres treated with proBDNF (100 ng/mL) showed dormancy without any neuronal migration and neurospheres had no morphological changes. Neurospheres treated with 2B11 (100 ng/ml) showed strong migration capability comparing with other groups. Neurospheres treated with 2B11 and proBDNF (100 ng/ml) showed similar ability of migration with sheep anti-proBDNF antibody treatment group.

Article Snippet: For ELISA assay, 96-well polystyrene microtiter plates (Nunc, Roskilde, Denmark, USA) were coated with the generated human proBDNF prodomain, and commercial human, rat and mice proBDNF proteins (Alomone Labs, Israel), and human mBDNF (Alomone Labs, Israel) (all at 1 μg/mL with total volume 50 μL) overnight at 4 °C.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Migration, Control, Comparison

(A) Representative images showing GFP and c-Fos (arrows) expression patterns in the PVT, a downstream target of the VMH, in AAV-DIO-EYFP, AAV-DIO-NaChBac+sgRNA, AAV-DIO-NaChBac+sgVglut2 injection SF1-Cre::ERa-Cre mice. scale bar=250 um. (B) Representative images showing GFP (green) and c-Fos (red, arrows) expression patterns in the PVT, a downstream target of the VMH, in SF1-Cre::ERa-Cre::BDNFflox/++EYFP, SF1-Cre::ERa-Cre::BDNFflox/++NaChBac, SF1-Cre::ERa-Cre::BDNFflox/flox+NaChBac mice. scale bar=250 um. (C) Number of c-Fos positive cells in the PVT region related to panel A. n=4-5 per group; one-way ANOVA; ***P<0.001 versus EYFP; ###P<0.001 versus NaChBac+sgRNA. (D) number of c-Fos positive cells in the PVT region related to panel A. n=4-5 per group; one-way ANOVA; ***P<0.001 versus EYFP; ###P<0.001 versus SF1-Cre::ERa-Cre:: BDNFflox/flox+NaChBac. (E) Diagram of viral delivery (AAV-DIO-mCherry, AAV-DIO-BDNF-mCherry+sgRNA, AAV-DIO-BDNF-mCherry+sgVglut2) into the VMH of SF1-Cre::ERa-Cre mice. (F) Body weight trajectories of SF1-Cre::ERa-Cre mice after VH injection of virus. n=4 (EYFP, BDNF+sgVglut2), n=5 (BDNF+sgRNA); two-way ANOVA; *P<0.05, **P<0.01, ***P<0.001 versus mCherry; #P<0.05 versus BDNF+sgRNA. (G) Daily food intake. n=4-5 per group, one-way ANOVA, **P=0.006 versus mCherry; #P=0.043 versus BDNF+sgRNA. (H) Total number of jumps during a 10-min test. n=4-5 per group; one-way ANOVA; ***P<0.01 versus mCherry; ###P<0.001 versus BDNF+sgRNA. (I, J) time spent in the light room in light-dark test (I), social time with stranger in three-chamber test (J). n=4-5 per group; one-way ANOVA; *P<0.05, ***P<0.01 versus mCherry; #P<0.05 versus BDNF+sgRNA. (K) Representative images showing mCherry (green) and c-Fos (magenta, arrows) expression patterns in the PVT, a downstream of VMH, in mCherry, BDNF+sgRNA, BDNF+sgVglut2 injection SF1-Cre::ERa-Cre mice. scale bar=250 um. (L) number of c-Fos positive cells in PVT region related to panel K. n=4-5 per group; one-way ANOVA; ***P<0.001 versus mCherry; ###P<0.001 versus BDNF+sgRNA.

Journal: bioRxiv

Article Title: Identification of a neural basis for anorexia nervosa

doi: 10.64898/2026.02.07.704578

Figure Lengend Snippet: (A) Representative images showing GFP and c-Fos (arrows) expression patterns in the PVT, a downstream target of the VMH, in AAV-DIO-EYFP, AAV-DIO-NaChBac+sgRNA, AAV-DIO-NaChBac+sgVglut2 injection SF1-Cre::ERa-Cre mice. scale bar=250 um. (B) Representative images showing GFP (green) and c-Fos (red, arrows) expression patterns in the PVT, a downstream target of the VMH, in SF1-Cre::ERa-Cre::BDNFflox/++EYFP, SF1-Cre::ERa-Cre::BDNFflox/++NaChBac, SF1-Cre::ERa-Cre::BDNFflox/flox+NaChBac mice. scale bar=250 um. (C) Number of c-Fos positive cells in the PVT region related to panel A. n=4-5 per group; one-way ANOVA; ***P<0.001 versus EYFP; ###P<0.001 versus NaChBac+sgRNA. (D) number of c-Fos positive cells in the PVT region related to panel A. n=4-5 per group; one-way ANOVA; ***P<0.001 versus EYFP; ###P<0.001 versus SF1-Cre::ERa-Cre:: BDNFflox/flox+NaChBac. (E) Diagram of viral delivery (AAV-DIO-mCherry, AAV-DIO-BDNF-mCherry+sgRNA, AAV-DIO-BDNF-mCherry+sgVglut2) into the VMH of SF1-Cre::ERa-Cre mice. (F) Body weight trajectories of SF1-Cre::ERa-Cre mice after VH injection of virus. n=4 (EYFP, BDNF+sgVglut2), n=5 (BDNF+sgRNA); two-way ANOVA; *P<0.05, **P<0.01, ***P<0.001 versus mCherry; #P<0.05 versus BDNF+sgRNA. (G) Daily food intake. n=4-5 per group, one-way ANOVA, **P=0.006 versus mCherry; #P=0.043 versus BDNF+sgRNA. (H) Total number of jumps during a 10-min test. n=4-5 per group; one-way ANOVA; ***P<0.01 versus mCherry; ###P<0.001 versus BDNF+sgRNA. (I, J) time spent in the light room in light-dark test (I), social time with stranger in three-chamber test (J). n=4-5 per group; one-way ANOVA; *P<0.05, ***P<0.01 versus mCherry; #P<0.05 versus BDNF+sgRNA. (K) Representative images showing mCherry (green) and c-Fos (magenta, arrows) expression patterns in the PVT, a downstream of VMH, in mCherry, BDNF+sgRNA, BDNF+sgVglut2 injection SF1-Cre::ERa-Cre mice. scale bar=250 um. (L) number of c-Fos positive cells in PVT region related to panel K. n=4-5 per group; one-way ANOVA; ***P<0.001 versus mCherry; ###P<0.001 versus BDNF+sgRNA.

Article Snippet: The following primary antibodies were used: rabbit anti-cFos (1:1000, #2250, Cell Signaling Technology), rabbit anti-BDNF(1:200, ANT-010, Alomone Labs), goat anti-GFP (1:1000, 51370, ROCKLAND), rabbit anti-ER alpha (1:500, NBP1-84827), chicken anti-mCherry (1:500, MCHERRY-0020, aves labs).The next day, sections were rinsed three times in PBS with 0.1% Triton X-100 and incubated with secondary antibodies prepared in the same diluent (secondary antibody, 5% normal donkey serum, 0.1% Triton X-100 in PBS).

Techniques: Expressing, Injection, Virus

Representative images showing mCherry (green) and c-Fos (magenta, arrows) expression pattern in lateral septum (A), preoptic area (C), PAG (E), periLC (G). scale bar=250 μm. The pictures in the bottom row of each panel represent the magnified pictures shown in the boxed area in each column. Number of cFos positive cells in in lateral septum (B), preoptic area (D), PAG (F, periLC (H). n=4-5 per group; one-way ANOVA; ***P<0.001 versus mCherry; ###P<0.001 versus BDNF+sgRNA.

Journal: bioRxiv

Article Title: Identification of a neural basis for anorexia nervosa

doi: 10.64898/2026.02.07.704578

Figure Lengend Snippet: Representative images showing mCherry (green) and c-Fos (magenta, arrows) expression pattern in lateral septum (A), preoptic area (C), PAG (E), periLC (G). scale bar=250 μm. The pictures in the bottom row of each panel represent the magnified pictures shown in the boxed area in each column. Number of cFos positive cells in in lateral septum (B), preoptic area (D), PAG (F, periLC (H). n=4-5 per group; one-way ANOVA; ***P<0.001 versus mCherry; ###P<0.001 versus BDNF+sgRNA.

Article Snippet: The following primary antibodies were used: rabbit anti-cFos (1:1000, #2250, Cell Signaling Technology), rabbit anti-BDNF(1:200, ANT-010, Alomone Labs), goat anti-GFP (1:1000, 51370, ROCKLAND), rabbit anti-ER alpha (1:500, NBP1-84827), chicken anti-mCherry (1:500, MCHERRY-0020, aves labs).The next day, sections were rinsed three times in PBS with 0.1% Triton X-100 and incubated with secondary antibodies prepared in the same diluent (secondary antibody, 5% normal donkey serum, 0.1% Triton X-100 in PBS).

Techniques: Expressing

Representative images showing mCherry (red) and cFos (cyan, arrows) expression pattern in VH in mCherry, BDNF+sgRNA, BDNF+sgVglut2 injection SF1-Cre::ERa-Cre mice (related to ). Scale bar=200 μm.

Journal: bioRxiv

Article Title: Identification of a neural basis for anorexia nervosa

doi: 10.64898/2026.02.07.704578

Figure Lengend Snippet: Representative images showing mCherry (red) and cFos (cyan, arrows) expression pattern in VH in mCherry, BDNF+sgRNA, BDNF+sgVglut2 injection SF1-Cre::ERa-Cre mice (related to ). Scale bar=200 μm.

Article Snippet: The following primary antibodies were used: rabbit anti-cFos (1:1000, #2250, Cell Signaling Technology), rabbit anti-BDNF(1:200, ANT-010, Alomone Labs), goat anti-GFP (1:1000, 51370, ROCKLAND), rabbit anti-ER alpha (1:500, NBP1-84827), chicken anti-mCherry (1:500, MCHERRY-0020, aves labs).The next day, sections were rinsed three times in PBS with 0.1% Triton X-100 and incubated with secondary antibodies prepared in the same diluent (secondary antibody, 5% normal donkey serum, 0.1% Triton X-100 in PBS).

Techniques: Expressing, Injection

FIGURE 1 (a) Workflow of data mining. The TMR (transmembrane sequence +5 amino acid residues from each N- and C-terminal sides) of curated entries found in UniProt using the code for tyrosine-kinase receptor family (2.7.10.1) were screened for the presence of cholesterol recognition and alignment consensus (CRAC and CARC). The incidence of CRAC and CARC motifs in (b) human, (c) mouse, (d) zebrafish, (e) fruit fly, and (f) C. elegans TMR of RTK family members. (g) Library of CRAC and CARC sequences (all combinations used can be found in the stored data). Residue sequence predicted to be embedded into the cell membrane shown in yellow

Journal: The European journal of neuroscience

Article Title: Cholesterol-recognition motifs in the transmembrane domain of the tyrosine kinase receptor family: The case of TRKB.

doi: 10.1111/ejn.15218

Figure Lengend Snippet: FIGURE 1 (a) Workflow of data mining. The TMR (transmembrane sequence +5 amino acid residues from each N- and C-terminal sides) of curated entries found in UniProt using the code for tyrosine-kinase receptor family (2.7.10.1) were screened for the presence of cholesterol recognition and alignment consensus (CRAC and CARC). The incidence of CRAC and CARC motifs in (b) human, (c) mouse, (d) zebrafish, (e) fruit fly, and (f) C. elegans TMR of RTK family members. (g) Library of CRAC and CARC sequences (all combinations used can be found in the stored data). Residue sequence predicted to be embedded into the cell membrane shown in yellow

Article Snippet: The plates were washed 3× with PBS buffer, and a mixture of biotinylated BDNF (bBDNF: 0, 0.05, 0.1, 0.5, 1, 5, or 10 ng/ml, Alomone Labs, #B- 250- B) and cholesterol (0, 20, 50 or 100 μM) was added for 1h at room temperature, followed by washing with PBS.

Techniques: Sequencing, Residue, Membrane

FIGURE 3 TRKB interaction with cholesterol. (a,b) Snapshots of the interaction between TRKB and cholesterol at the TMR (see supplement video) indicating that cholesterol (orange) interacts with the OH- group (red) in Y433 (cyan) of the TRKB.TMR (green). The TRKB.TMR helix is shown in the cartoon, the lipids and Y433 sidechain in van der Waals representations. (c) Molecular dynamics simulations show that cholesterol predominantly resides close to the C-alpha atom of Y433 but not C-alpha atom of R427, highlighting the significance of the aromatic Y in the TRKB CARC domain. (d) The binding of biotinylated BDNF (bBDNF) to TRKB is modulated by added cholesterol in a bell-shaped fashion [from Scatchard analysis Kd for bBDNF ctrl: 2.77 pM; chol 20 μM: 2.27pM; chol 50 μM: 2.61 pM; chol 100 μM: 1.93 pM; Bmax for bBDNF ctrl: 96.18; chol 20 μM: 108.9; chol 50 μM: 80.81; chol 100 μM: 65.35]. Curves from the Scatchard transformation of the TRKB:BDNF binding are depicted in the insert. Data expressed as mean/SEM of binding normalized by bBDNF10 ng/ml ctrl (no added cholesterol). (e) Expansion of the binding of bBDNF 1 ng/ml (black circles) and bBDNF 0 (ctrl, open circles), depicting the bell-shaped effect of different concentrations of cholesterol on bBDNF:TRKB interaction. Data expressed as mean/SEM of binding normalized by bBDNF 1 ng/ml ctrl. (f) Unlabeled BDNF (0-20 ng/ ml) dislocates bBDNF (1 ng/ml) binding to immobilized TRKB assayed at no supplemented cholesterol (0 μM). Data expressed as mean/SEM normalized by bBDNF 1 ng/ml at BDNF 0 ng/ml. *p < .05 from control (BDNF 1 ng/ml–cholesterol ctrl, R427)

Journal: The European journal of neuroscience

Article Title: Cholesterol-recognition motifs in the transmembrane domain of the tyrosine kinase receptor family: The case of TRKB.

doi: 10.1111/ejn.15218

Figure Lengend Snippet: FIGURE 3 TRKB interaction with cholesterol. (a,b) Snapshots of the interaction between TRKB and cholesterol at the TMR (see supplement video) indicating that cholesterol (orange) interacts with the OH- group (red) in Y433 (cyan) of the TRKB.TMR (green). The TRKB.TMR helix is shown in the cartoon, the lipids and Y433 sidechain in van der Waals representations. (c) Molecular dynamics simulations show that cholesterol predominantly resides close to the C-alpha atom of Y433 but not C-alpha atom of R427, highlighting the significance of the aromatic Y in the TRKB CARC domain. (d) The binding of biotinylated BDNF (bBDNF) to TRKB is modulated by added cholesterol in a bell-shaped fashion [from Scatchard analysis Kd for bBDNF ctrl: 2.77 pM; chol 20 μM: 2.27pM; chol 50 μM: 2.61 pM; chol 100 μM: 1.93 pM; Bmax for bBDNF ctrl: 96.18; chol 20 μM: 108.9; chol 50 μM: 80.81; chol 100 μM: 65.35]. Curves from the Scatchard transformation of the TRKB:BDNF binding are depicted in the insert. Data expressed as mean/SEM of binding normalized by bBDNF10 ng/ml ctrl (no added cholesterol). (e) Expansion of the binding of bBDNF 1 ng/ml (black circles) and bBDNF 0 (ctrl, open circles), depicting the bell-shaped effect of different concentrations of cholesterol on bBDNF:TRKB interaction. Data expressed as mean/SEM of binding normalized by bBDNF 1 ng/ml ctrl. (f) Unlabeled BDNF (0-20 ng/ ml) dislocates bBDNF (1 ng/ml) binding to immobilized TRKB assayed at no supplemented cholesterol (0 μM). Data expressed as mean/SEM normalized by bBDNF 1 ng/ml at BDNF 0 ng/ml. *p < .05 from control (BDNF 1 ng/ml–cholesterol ctrl, R427)

Article Snippet: The plates were washed 3× with PBS buffer, and a mixture of biotinylated BDNF (bBDNF: 0, 0.05, 0.1, 0.5, 1, 5, or 10 ng/ml, Alomone Labs, #B- 250- B) and cholesterol (0, 20, 50 or 100 μM) was added for 1h at room temperature, followed by washing with PBS.

Techniques: Binding Assay, Transformation Assay, Control

Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the prefrontal (prelimbic/infralimbic) cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. ***: p < 0.001 (post hoc Duncan’s multiple range test).

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the prefrontal (prelimbic/infralimbic) cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. ***: p < 0.001 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the anterior cingulate cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. **: p < 0.01 (post hoc Duncan’s multiple range test).

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the anterior cingulate cortex of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. **: p < 0.01 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the ventral hippocampus of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. *: p < 0.05; **: p < 0.02 (post hoc Duncan’s multiple range test).

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the ventral hippocampus of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group. *: p < 0.05; **: p < 0.02 (post hoc Duncan’s multiple range test).

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the dorsal hippocampus of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group.

Journal: Brain Sciences

Article Title: Neonatal Handling Positively Modulates Anxiety, Sensorimotor Gating, Working Memory, and Cortico-Hippocampal Neuroplastic Adaptations in Two Genetically Selected Rat Strains Differing in Emotional and Cognitive Traits

doi: 10.3390/brainsci15080776

Figure Lengend Snippet: Western blot analysis of the BDNF ( A , B ), trkB ( C , D ), and the PSA-NCAM ( E , F ), in the dorsal hippocampus of the RHA and the RLA rats, either untreated controls (CTRL) or treated with neonatal handling (NH). The values represent the densitometric analysis of the BDNF/GAPDH ( B ), trkB/GAPDH ( D ), and the PSA-NCAM/GAPDH ( F ) band grey optical density (O.D.) ratios. The bars denote the mean ± S.E.M. of 9–10 rats, in each experimental group.

Article Snippet: Molecular weight (mw) standards (Precision Plus Protein Western C Standards, Cat# 161–0376, Bio-Rad, Hercules, CA, USA) and recombinant human BDNF protein (rhBDNF) (Cat# B-257, Alomone Labs, Jerusalem, Israel) were run in parallel.

Techniques: Western Blot

Fig. 3. Suppression of treRNA by miR-190a inhibits EMT in HCC. (A) 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or sitreRNA2), 20 nM miR-190a, or NC RNA was transfected into Huh-7 or HepG2 cells. EMT related genes were quantified 72 h post-transfection as indicated. (B) Cells were treated with antagomiR (anti-miR- 190a) or NC RNA as in (A). The mRNA level of different EMT genes was quantified and normalized to b-actin. (C) Cell lysates were blotting with EMT marker E-cadherin (E-cad), Claudin-1 (Claud), Vimentin (Vimen), N-cadherin (N-cad), SNAI1 (Snail). b-Actin was used as endogenous control. The EMT marker expression level in was calculated using Image J and presented as histogram (right panel). (D) Cells were treated with TGF-b for 12 h and transfected with 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or si-treRNA2) or NC RNA subsequently. The morphological change was captured 24 h after transfection.

Journal: FEBS letters

Article Title: miR-190a inhibits epithelial-mesenchymal transition of hepatoma cells via targeting the long non-coding RNA treRNA.

doi: 10.1016/j.febslet.2015.11.024

Figure Lengend Snippet: Fig. 3. Suppression of treRNA by miR-190a inhibits EMT in HCC. (A) 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or sitreRNA2), 20 nM miR-190a, or NC RNA was transfected into Huh-7 or HepG2 cells. EMT related genes were quantified 72 h post-transfection as indicated. (B) Cells were treated with antagomiR (anti-miR- 190a) or NC RNA as in (A). The mRNA level of different EMT genes was quantified and normalized to b-actin. (C) Cell lysates were blotting with EMT marker E-cadherin (E-cad), Claudin-1 (Claud), Vimentin (Vimen), N-cadherin (N-cad), SNAI1 (Snail). b-Actin was used as endogenous control. The EMT marker expression level in was calculated using Image J and presented as histogram (right panel). (D) Cells were treated with TGF-b for 12 h and transfected with 20 nM miR-190a, 20 nM siRNA duplexes against treRNA (si-treRNA1 or si-treRNA2) or NC RNA subsequently. The morphological change was captured 24 h after transfection.

Article Snippet: Membranes were blocked by 5% BSA or 5% non-fat milk for 3 h at 25 C and incubated with primary antibody against Ago2 (#2897, Cell Signaling, US), Dicer (#5362, Cell Signaling), E-Cadherin (#3195, Cell Signaling), Claudin-1 (#13255, Cell Signaling), Vimentin (#5741, Cell Signaling), N-Cadherin (#13116, Cell Signaling), SNAI1 (#3897, Cell Signaling), or b-actin (C-4, Santa cruz, US) for 12 h at 4 C. Membranes were incubated with secondary antibody diluted in Tris-buffered saline supplemented with 0.5% Tween-20 and 5% BSA or 5% non-fat milk for 1 h at 25 C. The signal was detected with the enhanced chemiluminescence system (Pierce, US) and exposed to X-ray film (Kodak, US).

Techniques: Transfection, Marker, Control, Expressing

Cellular localization and uptake of fluorescently labeled sEVs with SCs. (A) Cellular colocalization: SP8 was used to photograph sEVs prelabeled with PKH67 (green fluorescence) with Hoechst 33,342 (blue fluorescence)-stained SC nuclei. (B) Western blot: Detection of ERK1/2, ZEB2, and c-JUN expression levels in NC Schwann cells and after 48-h treatment with hypoxia sEVs. (C) Real-time PCR: Inflammatory and other restoration-related factors (IL-1β, IL-6, TNF-α, etc.) were detected. (D) ELISA test: GDNF NDF and NT-3 in the supernatant of Schwann cell preparation. Statistical significance, * p < 0.05,** p < 0.01, and **** p < 0.0001. *Significant difference.

Journal: Frontiers in Cellular Neuroscience

Article Title: Hypoxic culture of umbilical cord mesenchymal stem cell-derived sEVs prompts peripheral nerve injury repair

doi: 10.3389/fncel.2022.897224

Figure Lengend Snippet: Cellular localization and uptake of fluorescently labeled sEVs with SCs. (A) Cellular colocalization: SP8 was used to photograph sEVs prelabeled with PKH67 (green fluorescence) with Hoechst 33,342 (blue fluorescence)-stained SC nuclei. (B) Western blot: Detection of ERK1/2, ZEB2, and c-JUN expression levels in NC Schwann cells and after 48-h treatment with hypoxia sEVs. (C) Real-time PCR: Inflammatory and other restoration-related factors (IL-1β, IL-6, TNF-α, etc.) were detected. (D) ELISA test: GDNF NDF and NT-3 in the supernatant of Schwann cell preparation. Statistical significance, * p < 0.05,** p < 0.01, and **** p < 0.0001. *Significant difference.

Article Snippet: The following ELISA kits were: mouse glial cell line-derived neurotrophic factor (GDNF), ELISA kit (CSB-E07341m, CUSABIO), mouse neurotrophin 3 (NT-3), ELISA kit (CSB-E04687m, CUSABIO), and mouse nerve growth factor (NGF), ELISA kit (CSB-E04684m, CUSABIO).

Techniques: Labeling, Fluorescence, Staining, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay