Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: JKZP‐containing serum may enhance the synaptic plasticity of hippocampal neurons by interfering with S100A10/tPA and modulating the proBDNF/mBDNF balance. (A) Neuronal growth process (×200); (B) MAP2 immunofluorescence to identify the purity of neurons (the purity was >95%, ×200, n = 5); (C) CCK8 screening of neurons for OGD modeling time ( n = 6); (D) CCK8 detection of the toxic effects of JKZP‐containing serum lyophilized powder on neurons ( n = 5); (E) CCK8 detection of the effects of JKZP‐containing serum lyophilized powder on neuronal activity ( n = 5). (F) RT‐qPCR assay for PSD95, GAP43, and SYN mRNA expressions ( n = 3); (G) representative images of IF staining for SYN, GAP43, and PSD95 and corresponding quantitative analyses ( n = 4, ×200); (H) WB assay for PSD95, GAP43, and SYN protein expressions ( n = 3); (I) WB assay for S100A10/tPA/BDNF pathway related protein levels ( n = 3). * p < 0.05, ** p < 0.01. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; P75 NTR , P75 neurotrophic receptor; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator; TrKB, tropomyosin receptor kinase B.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Immunofluorescence, Activity Assay, Quantitative RT-PCR, Staining, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: Primer sequences.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques:

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: Effects of JKZP on S100A10/tPA/BDNF pathway. (A–D) Representative images of IF staining for S100A10 and tPA, and corresponding quantitative analyses (×1000); (E) RT‐qPCR analysis of detection of mBDNF, TrkB, and P75 NTR mRNA expression; (F and G) WB analysis of BDNF pathway protein expression. * p < 0.05, ** p < 0.01. n = 3. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; P75NTR, P75 neurotrophic receptor; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator; TrKB, tropomyosin receptor kinase B.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Staining, Quantitative RT-PCR, Expressing, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: JKZP‐containing serum may enhance the synaptic plasticity of hippocampal neurons by interfering with S100A10/tPA and modulating the proBDNF/mBDNF balance. (A) Neuronal growth process (×200); (B) MAP2 immunofluorescence to identify the purity of neurons (the purity was >95%, ×200, n = 5); (C) CCK8 screening of neurons for OGD modeling time ( n = 6); (D) CCK8 detection of the toxic effects of JKZP‐containing serum lyophilized powder on neurons ( n = 5); (E) CCK8 detection of the effects of JKZP‐containing serum lyophilized powder on neuronal activity ( n = 5). (F) RT‐qPCR assay for PSD95, GAP43, and SYN mRNA expressions ( n = 3); (G) representative images of IF staining for SYN, GAP43, and PSD95 and corresponding quantitative analyses ( n = 4, ×200); (H) WB assay for PSD95, GAP43, and SYN protein expressions ( n = 3); (I) WB assay for S100A10/tPA/BDNF pathway related protein levels ( n = 3). * p < 0.05, ** p < 0.01. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; P75 NTR , P75 neurotrophic receptor; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator; TrKB, tropomyosin receptor kinase B.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Immunofluorescence, Activity Assay, Quantitative RT-PCR, Staining, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: Primer sequences.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques:

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: JKZP improves neuronal synaptic plasticity by regulating the balance of proBDNF/mBDNF through S100A10/tPA. (A) Fluorescence microscopy to observe the transfection rate (×200); (B) RT‐qPCR to detect the cellular S100A10 knockdown rate ( n = 4); (C and D) representative images of IF staining for SYN, GAP43, and PSD95 and corresponding quantitative analyses ( n = 4, ×200); (E) WB detection of synapse‐associated protein expression ( n = 4); (F) WB assay for S100A10/tPA/BDNF pathway related protein expression ( n = 4); (G) ELISA detection of tPA protein expression ( n = 6). * p < 0.05, ** p < 0.01. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; MOI, multiplicity of infection; OGD/R, oxygen‐glucose deprivation/reperfusion; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Fluorescence, Microscopy, Transfection, Quantitative RT-PCR, Knockdown, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Infection, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: Effects of JKZP on S100A10/tPA/BDNF pathway. (A–D) Representative images of IF staining for S100A10 and tPA, and corresponding quantitative analyses (×1000); (E) RT‐qPCR analysis of detection of mBDNF, TrkB, and P75 NTR mRNA expression; (F and G) WB analysis of BDNF pathway protein expression. * p < 0.05, ** p < 0.01. n = 3. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; P75NTR, P75 neurotrophic receptor; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator; TrKB, tropomyosin receptor kinase B.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Staining, Quantitative RT-PCR, Expressing, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: JKZP‐containing serum may enhance the synaptic plasticity of hippocampal neurons by interfering with S100A10/tPA and modulating the proBDNF/mBDNF balance. (A) Neuronal growth process (×200); (B) MAP2 immunofluorescence to identify the purity of neurons (the purity was >95%, ×200, n = 5); (C) CCK8 screening of neurons for OGD modeling time ( n = 6); (D) CCK8 detection of the toxic effects of JKZP‐containing serum lyophilized powder on neurons ( n = 5); (E) CCK8 detection of the effects of JKZP‐containing serum lyophilized powder on neuronal activity ( n = 5). (F) RT‐qPCR assay for PSD95, GAP43, and SYN mRNA expressions ( n = 3); (G) representative images of IF staining for SYN, GAP43, and PSD95 and corresponding quantitative analyses ( n = 4, ×200); (H) WB assay for PSD95, GAP43, and SYN protein expressions ( n = 3); (I) WB assay for S100A10/tPA/BDNF pathway related protein levels ( n = 3). * p < 0.05, ** p < 0.01. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; P75 NTR , P75 neurotrophic receptor; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator; TrKB, tropomyosin receptor kinase B.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Immunofluorescence, Activity Assay, Quantitative RT-PCR, Staining, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: JKZP improves neuronal synaptic plasticity by regulating the balance of proBDNF/mBDNF through S100A10/tPA. (A) Fluorescence microscopy to observe the transfection rate (×200); (B) RT‐qPCR to detect the cellular S100A10 knockdown rate ( n = 4); (C and D) representative images of IF staining for SYN, GAP43, and PSD95 and corresponding quantitative analyses ( n = 4, ×200); (E) WB detection of synapse‐associated protein expression ( n = 4); (F) WB assay for S100A10/tPA/BDNF pathway related protein expression ( n = 4); (G) ELISA detection of tPA protein expression ( n = 6). * p < 0.05, ** p < 0.01. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; MOI, multiplicity of infection; OGD/R, oxygen‐glucose deprivation/reperfusion; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Fluorescence, Microscopy, Transfection, Quantitative RT-PCR, Knockdown, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Infection, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: Effects of JKZP on S100A10/tPA/BDNF pathway. (A–D) Representative images of IF staining for S100A10 and tPA, and corresponding quantitative analyses (×1000); (E) RT‐qPCR analysis of detection of mBDNF, TrkB, and P75 NTR mRNA expression; (F and G) WB analysis of BDNF pathway protein expression. * p < 0.05, ** p < 0.01. n = 3. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; P75NTR, P75 neurotrophic receptor; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator; TrKB, tropomyosin receptor kinase B.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Staining, Quantitative RT-PCR, Expressing, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: JKZP‐containing serum may enhance the synaptic plasticity of hippocampal neurons by interfering with S100A10/tPA and modulating the proBDNF/mBDNF balance. (A) Neuronal growth process (×200); (B) MAP2 immunofluorescence to identify the purity of neurons (the purity was >95%, ×200, n = 5); (C) CCK8 screening of neurons for OGD modeling time ( n = 6); (D) CCK8 detection of the toxic effects of JKZP‐containing serum lyophilized powder on neurons ( n = 5); (E) CCK8 detection of the effects of JKZP‐containing serum lyophilized powder on neuronal activity ( n = 5). (F) RT‐qPCR assay for PSD95, GAP43, and SYN mRNA expressions ( n = 3); (G) representative images of IF staining for SYN, GAP43, and PSD95 and corresponding quantitative analyses ( n = 4, ×200); (H) WB assay for PSD95, GAP43, and SYN protein expressions ( n = 3); (I) WB assay for S100A10/tPA/BDNF pathway related protein levels ( n = 3). * p < 0.05, ** p < 0.01. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; P75 NTR , P75 neurotrophic receptor; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator; TrKB, tropomyosin receptor kinase B.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Immunofluorescence, Activity Assay, Quantitative RT-PCR, Staining, Binding Assay

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: Primer sequences.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques:

Journal: Brain and Behavior

Article Title: Jiawei Kongsheng Zhenzhong Pill (JKZP) Alleviates Chronic Cerebral Hypoperfusion‐Induced Hippocampal Synaptic Damage via S100A10/tPA/BDNF Pathway

doi: 10.1002/brb3.70328

Figure Lengend Snippet: Effects of JKZP on S100A10/tPA/BDNF pathway. (A–D) Representative images of IF staining for S100A10 and tPA, and corresponding quantitative analyses (×1000); (E) RT‐qPCR analysis of detection of mBDNF, TrkB, and P75 NTR mRNA expression; (F and G) WB analysis of BDNF pathway protein expression. * p < 0.05, ** p < 0.01. n = 3. JKZP, Jiawei Kongsheng Zhenzhong Pill ; mBDNF, mature BDNF; P75NTR, P75 neurotrophic receptor; proBDNF, protein of BDNF; S100A10, S100 calcium‐binding protein A10; tPA, tissue‐type plasminogen activator; TrKB, tropomyosin receptor kinase B.

Article Snippet: Primary antibodies including growth‐associated protein 43 (GAP43, Cat. ab75810), synaptophysin (SYN, Cat. ab32127), Neun (Cat. ab177487, Abcam), MAP2 (ab254264, Abcam), and GAPDH (Cat. ab8245, Abcam) were purchased from Abcam, USA; postsynaptic density protein 95 (PSD95, Cat. 12369‐1‐AP), tPA (Cat. 10147‐1‐AP), BDNF (Cat. 28205‐1‐AP), TrkB (Cat. 13129‐1‐AP), P75 NTR (Cat. 55014‐1‐AP), sortilin (Cat. 12369‐1‐AP), and β‐actin (Cat. HRP‐60008) were purchased from Proteintech Group Inc., China; S100A10 (Cat. 201226) was purchased from Zen‐Bio, China.

Techniques: Staining, Quantitative RT-PCR, Expressing, Binding Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Involvement of neurotrophic signaling in doxorubicin-induced cardiotoxicity

doi: 10.3892/etm.2019.8276

Figure Lengend Snippet: Effects of DOX on the expression of BDNF and NGF in the serum and heart. (A) BDNF concentration in the serum. (B) BDNF mRNA levels in the heart. (C) BDNF protein expression levels in the heart. (D) NGF concentration in the serum. (E) NGF mRNA levels in the heart. (F) NGF protein levels in the heart. Data are expressed as the mean ± standard deviation (n=8–9). *P<0.05 and **P<0.01 compared with Con. NGF, nerve growth factor; BDNF, brain derived neurotrophic factor; DOX, doxorubicin; Con, control.

Article Snippet: Blots were incubated at 4°C overnight with antibodies against BDNF (Abcam; cat. no. ab1083191; 1:2,000); NGF (Abcam; cat. no. ab6199; 1:1,000); TrkA (Abcam; cat. no. ab76291; 1:3,000); p-TrkA (Tyr49; Cell Signaling Technology, Inc.; cat. no. 9141; 1:1,000); TrkB (Santa Cruz Biotechnology, Inc.; cat. no. sc377218; 1:200); p-TrkB (Tyr705; Thermo Fisher Scientific, Inc.; cat. no. PA538077; 1:500); p75NTR (ProteinTech Group, Inc.; cat. no. 55014-1-AP; 1:1,000); Akt (ProteinTech Group, Inc.; cat. no. 10176-2-AP; 1:500); pAkt (Ser473; Cell Signaling Technology, Inc.; cat. no. 4060; 1:3,000); Erk (Santa Cruz Biotechnology, Inc.; cat. no. sc514302; 1:200); pErk (Thr202/Tyr204; Cell Signaling Technology, Inc.; cat. no. 4695; 1:2,000) and β-actin (ProteinTech Group, Inc.; cat. no. 66009-1-Ig; 1:4,000).

Techniques: Expressing, Concentration Assay, Standard Deviation, Derivative Assay

Journal: Neural Regeneration Research

Article Title: Small extracellular vesicles derived from hair follicle neural crest stem cells enhance perineurial cell proliferation and migration via the TGF-β/SMAD/HAS2 pathway

doi: 10.4103/NRR.NRR-D-25-00127

Figure Lengend Snippet: hfNCSC-sEVs enhance tube formation and barrier function in PCs and promote tight junction protein expression. (A) Optical micrographs of the tube formation assay and (B) statistical analyses demonstrated the number of junctions and total length of tubes in PCs in both the phosphate-buffered saline (PBS) and hfNCSC-sEVs groups ( n = 5 per group). (C) Measurements of transmembrane resistance ( n = 3 per group) and (D) cell monolayer permeability assays ( n = 9 per group) indicated the barrier formation ability of PCs in both the PBS and hfNCSC-sEVs groups. (E) Western blot and (F) statistical analyses revealed the relative protein expression levels of the tight junction proteins zonula occludens 1 (ZO1) and claudin-1 in PCs from the PBS and hfNCSC-sEVs groups on day 7 of in vitro culture (normalized to β-actin, n = 3 per group). (G, H) Immunofluorescence staining (G) and statistical analyses (H) showed the integrated optical density (IOD) of ZO1 (green) and the expression of β-tubulin (red) in PCs from the PBS and hfNCSC-sEVs groups on day 7 of in vitro culture ( n = 3 per group). (I) Schematic illustration of the rat sciatic nerve defect model: a 5-mm defect was surgically created in the rat sciatic nerve, which was then bridged using a silicon tube, followed by an orthotopic injection procedure. (J) Immunofluorescence staining revealed the expression of claudin-1 (red) in the proximal end of regenerated tissue in both the PBS and hfNCSC-sEVs groups on day 7 post-operation, with 4′,6-diamidino-2-phenylindole (DAPI) staining indicating the nuclei. Data are expressed as the mean ± SEM. * P < 0.05, *** P < 0.001 (Student’s t -test for B, C, D, F, and H). The data were from at least three separate and independent studies. hfNCSCs: Hair follicle neural crest stem cells; IOD: integrated optical density; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-p75 neurotrophin receptor (p75) antibody (1:100, Cat# 55014-1-AP, Proteintech), mouse monoclonal anti-nestin antibody (1:100, Cat# MAB353, Sigma), rabbit polyclonal anti-claudin-1 antibody (1:250, Cat# 13050-1-AP, Proteintech), rabbit polyclonal anti-ZO1 antibody (1:200, Cat# 21773-1-AP, Proteintech), rabbit polyclonal anti-glucose transporter 1 (GLUT1) antibody (1:500, Cat# 21829-1-AP, Proteintech), rabbit monoclonal anti-S100 antibody (1:800, Cat# MAB353, Abcam), mouse monoclonal anti-neurofilament 200 (NF200) antibody (1:800, Cat# N5389, Sigma), rabbit polyclonal anti-myelin basic protein (MBP) antibody (1:400, Cat# 10458-1-AP, Proteintech), mouse monoclonal anti-β-tubulin antibody (1:1000, Cat# M20005 , Abmart), and rabbit polyclonal anti-HAS2 antibody (1:200, Cat# DF13702, Affinity).

Techniques: Expressing, Tube Formation Assay, Saline, Permeability, Western Blot, In Vitro, Immunofluorescence, Staining, Injection

Journal: Neural Regeneration Research

Article Title: Small extracellular vesicles derived from hair follicle neural crest stem cells enhance perineurial cell proliferation and migration via the TGF-β/SMAD/HAS2 pathway

doi: 10.4103/NRR.NRR-D-25-00127

Figure Lengend Snippet: miR-21-5p in hfNCSC-sEVs augments cell proliferation and migration by enhancing HAS2 expression in PCs. (A, B) Western blot (A) and statistical analyses (B) revealed the relative protein expression levels of HAS2, proliferating cell nuclear antigen (PCNA), and vimentin in PCs across the –/–, –/si- Has2 , hfNCSC-sEVs/–, and hfNCSC-sEVs/si- Has2 groups on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). (C, D) The wound healing assay (C) and statistical analysis (D) demonstrated the migration rates of PCs in the aforementioned groups ( n = 3 per group). (E) The Cell Counting Kit-8 assay was used to assess cell viability in PCs across the same groups on day 5 of in vitro culture ( n = 5 per group). (F, G) Western blot (F) and statistical analyses (G) indicated the relative protein expression levels of HAS2, PCNA, and vimentin in PCs treated with phosphate-buffered saline (PBS), hfNCSC-sEVs, or hfNCSC-sEVs + miR-21-5p inhibitor on day 5 of in vitro culture (normalized to β-actin, n = 3 per group). (H–J) Immunofluorescence staining visualized the expression of HAS2 (red) and 5-ethynyl-2′-deoxyuridine (EdU; green) in PCs (H), and statistical analysis revealed the integrated optical density (IOD) of zonula occludens 1 (ZO1; I) and the cell proliferation rates (J) in the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 5 of in vitro culture ( n = 3 per group). (K, L) Western blot (K) and statistical analyses (L) showed the relative protein expression levels of HAS2, PCNA, and vimentin in regenerated tissue from the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 5 post-operation (normalized to β-tubulin, n = 3 per group). Data are expressed as the mean ± SEM. ** P < 0.01, *** P < 0.001 (one-way analysis of variance and Tukey’s multiple comparison test for B, D, E, G, I, J, and L). The data were from at least three separate and independent studies. CCK-8: Cell counting kit-8; EdU: 5-ethynyl-2′-deoxyuridine; HAS2: hyaluronan synthase 2; hfNCSCs: hair follicle neural crest stem cells; IOD: integrated optical density; PCNA: proliferating cell nuclear antigen; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-p75 neurotrophin receptor (p75) antibody (1:100, Cat# 55014-1-AP, Proteintech), mouse monoclonal anti-nestin antibody (1:100, Cat# MAB353, Sigma), rabbit polyclonal anti-claudin-1 antibody (1:250, Cat# 13050-1-AP, Proteintech), rabbit polyclonal anti-ZO1 antibody (1:200, Cat# 21773-1-AP, Proteintech), rabbit polyclonal anti-glucose transporter 1 (GLUT1) antibody (1:500, Cat# 21829-1-AP, Proteintech), rabbit monoclonal anti-S100 antibody (1:800, Cat# MAB353, Abcam), mouse monoclonal anti-neurofilament 200 (NF200) antibody (1:800, Cat# N5389, Sigma), rabbit polyclonal anti-myelin basic protein (MBP) antibody (1:400, Cat# 10458-1-AP, Proteintech), mouse monoclonal anti-β-tubulin antibody (1:1000, Cat# M20005 , Abmart), and rabbit polyclonal anti-HAS2 antibody (1:200, Cat# DF13702, Affinity).

Techniques: Migration, Expressing, Western Blot, In Vitro, Wound Healing Assay, Cell Counting, Saline, Immunofluorescence, Staining, Comparison, CCK-8 Assay

Journal: Neural Regeneration Research

Article Title: Small extracellular vesicles derived from hair follicle neural crest stem cells enhance perineurial cell proliferation and migration via the TGF-β/SMAD/HAS2 pathway

doi: 10.4103/NRR.NRR-D-25-00127

Figure Lengend Snippet: miR-21-5p in hfNCSC-sEVs enhances tight junction protein expression in PCs. (A, B) Immunofluorescence staining (A) and statistical analysis (B) demonstrated IOD of ZO1 (green) and the expression of β-tubulin (red) in PCs across the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 of in vitro culture ( n = 3 per group). (C) Western blot and (D) statistical analyses revealed the relative protein expression levels of the tight junction proteins ZO1 and claudin-1 in PCs from the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 of in vitro culture (normalized to β-actin, n = 3 per group). (E) Immunofluorescence staining depicted the expression of claudin-1 (red) at the proximal end of regenerated tissue in the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 post-operation, with DAPI staining highlighting the nuclei. (F, G) Western blot (F) and statistical analyses (G) indicated the relative protein expression levels of ZO1 and claudin-1 in regenerated tissue across the PBS, hfNCSC-sEVs, and hfNCSC-sEVs + miR-21-5p inhibitor groups on day 7 post-operation (normalized to β-actin, n = 3 per group). Data are expressed as the mean ± SEM. ** P < 0.01, *** P < 0.001 (one-way analysis of variance and Tukey’s multiple comparison test for B, D, and G). The data were from at least three separate and independent studies. DAPI: 4,6-Diamidino-2-phenylindole; hfNCSCs: hair follicle neural crest stem cells; IOD: integrated optical density; PBS: phosphate-buffered saline; PCs: perineurial cells; sEVs: small extracellular vesicles; ZO1: zonula occludens 1.

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-p75 neurotrophin receptor (p75) antibody (1:100, Cat# 55014-1-AP, Proteintech), mouse monoclonal anti-nestin antibody (1:100, Cat# MAB353, Sigma), rabbit polyclonal anti-claudin-1 antibody (1:250, Cat# 13050-1-AP, Proteintech), rabbit polyclonal anti-ZO1 antibody (1:200, Cat# 21773-1-AP, Proteintech), rabbit polyclonal anti-glucose transporter 1 (GLUT1) antibody (1:500, Cat# 21829-1-AP, Proteintech), rabbit monoclonal anti-S100 antibody (1:800, Cat# MAB353, Abcam), mouse monoclonal anti-neurofilament 200 (NF200) antibody (1:800, Cat# N5389, Sigma), rabbit polyclonal anti-myelin basic protein (MBP) antibody (1:400, Cat# 10458-1-AP, Proteintech), mouse monoclonal anti-β-tubulin antibody (1:1000, Cat# M20005 , Abmart), and rabbit polyclonal anti-HAS2 antibody (1:200, Cat# DF13702, Affinity).

Techniques: Expressing, Immunofluorescence, Staining, In Vitro, Western Blot, Comparison, Saline