mann-whitney graphpad prism version 5.00 mac Search Results


r studio software version 0.99.483  (RStudio)
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RStudio r studio software version 0.99.483
R Studio Software Version 0.99.483, supplied by RStudio, 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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ht7800  (Hitachi Ltd)
99
Hitachi Ltd ht7800
Ht7800, supplied by Hitachi Ltd, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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version 15  (MedCalc Software Ltd)
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MedCalc Software Ltd version 15
Version 15, supplied by MedCalc Software Ltd, 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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sir-tubulin  (Spirochrome)
96
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drug medetomidine pfizer animal health  (Pfizer Animal Health)
86
Pfizer Animal Health drug medetomidine pfizer animal health
Figure 8. Effect of selective inhibition of pre-Bötzinger complex (preBötC)→7n neurons on mystacial pad (MP) activity is state-dependent. Schematic diagrams showing the injection protocols for non-selective transduction of preBötC neurons (A) and selective transduction of preBötC→7n neurons (K). Representative trace showing the integrated (∫) and raw MP EMG, heart rate (HR), electrocardiograms (ECG), and ∫ and raw diaphragm (Dia) EMG in the initial phase of reversal of <t>ketamine/medetomidine</t> anesthesia in rats with non-selective (B) and selective (L) preBötC neuron transduction. The period of photoinhibition is highlighted with the blue box. Group data showing mean (solid line) and 95% confidence intervals for (C and M) respiratory frequency - fR, (E and O) Dia amplitude, (G and Q) and HR (bpm) before, during, and after photoinhibition in non-selective (red), selective GtACR2 expressing (blue), and control (black) rats. Histograms showing group data for the effect of photoinhibition of preBötC on respiratory and cardiovascular parameters (D and N) fR, (F and P) Dia amplitude, (H and R) HR, (I and S) apnea duration and number of missed respiratory cycles, and (J and T) MP activity.; P1 and P2 refer to the initial period of photoinhibition. Group data are presented as mean ± 95% CI; unpaired t-test or nonparametric Mann- Whitney test with multiple comparisons using the Bonferroni-Dunn method, *p<0.05. The photoinhibition of preBötC neurons is depicted by blue
Drug Medetomidine Pfizer Animal Health, supplied by Pfizer Animal Health, 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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antibody foxo1  (Proteintech)
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Proteintech antibody foxo1
Figure 4. Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the <t>PI3K/AKT/FOXO1</t> pathway. Effects of US-CM and GDNF on the phosphorylated FOXO1 (p-FOXO1)/FOXO1 ratio, total FOXO1 expression (A), cytoplasmic p-FOXO1, cytoplasmic FOXO1, and nuclear FOXO1 expression (B). The expression levels of total and nuclear FOXO1 (C), claudin-5, and VE-cadherin (D) in hCMEC/D3 cells transfected with FOXO1 siRNA (siFOXO1). NC: negative control. (E) Effects of FOXO1 overexpression (FOXO1-OE) and GDNF on the expression levels of claudin-5, total FOXO1, and nuclear FOXO1. FOXO1-NC: negative control plasmids. (F) Effects of LY and U0 on GDNF- induced alterations of total p-FOXO1/FOXO1 ratio, cytoplasmic p-FOXO1, cytoplasmic FOXO1, and nuclear FOXO1 expression. (G) Effects of LY on the claudin-5 expression upregulated by siFOXO1. The above data are shown as the mean± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test, Welch’s ANOVA test, or Kruskal– Wallis test.
Antibody Foxo1, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibody ets1  (Proteintech)
94
Proteintech antibody ets1
Figure 5. Glial cell line-derived neurotrophic factor (GDNF)-induced VE-cadherin expression in hCMEC/D3 cells by activating the <t>PI3K/AKT/ETS1</t> and MAPK/ERK/ETS1 pathways. Effects of US-CM and GDNF on total (A) and nuclear (B) ETS1 expression. Effects of LY and U0 on 200 pg/ml GDNF-induced total (C) and nuclear (D) ETS1 expression. Expression levels of total (E) and the nuclear ETS1 (F) in hCMEC/D3 cells after knocking down ETS1 with siRNA (siETS1). (G) Effects of GDNF and siETS1 on the expression of VE-cadherin and claudin-5. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test.
Antibody Ets1, supplied by Proteintech, 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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Image Search Results


Figure 8. Effect of selective inhibition of pre-Bötzinger complex (preBötC)→7n neurons on mystacial pad (MP) activity is state-dependent. Schematic diagrams showing the injection protocols for non-selective transduction of preBötC neurons (A) and selective transduction of preBötC→7n neurons (K). Representative trace showing the integrated (∫) and raw MP EMG, heart rate (HR), electrocardiograms (ECG), and ∫ and raw diaphragm (Dia) EMG in the initial phase of reversal of ketamine/medetomidine anesthesia in rats with non-selective (B) and selective (L) preBötC neuron transduction. The period of photoinhibition is highlighted with the blue box. Group data showing mean (solid line) and 95% confidence intervals for (C and M) respiratory frequency - fR, (E and O) Dia amplitude, (G and Q) and HR (bpm) before, during, and after photoinhibition in non-selective (red), selective GtACR2 expressing (blue), and control (black) rats. Histograms showing group data for the effect of photoinhibition of preBötC on respiratory and cardiovascular parameters (D and N) fR, (F and P) Dia amplitude, (H and R) HR, (I and S) apnea duration and number of missed respiratory cycles, and (J and T) MP activity.; P1 and P2 refer to the initial period of photoinhibition. Group data are presented as mean ± 95% CI; unpaired t-test or nonparametric Mann- Whitney test with multiple comparisons using the Bonferroni-Dunn method, *p<0.05. The photoinhibition of preBötC neurons is depicted by blue

Journal: eLife

Article Title: Selective transduction and photoinhibition of pre-Bötzinger neurons that project to the facial nucleus in rats affect the nasofacial activity

doi: 10.7554/elife.85398

Figure Lengend Snippet: Figure 8. Effect of selective inhibition of pre-Bötzinger complex (preBötC)→7n neurons on mystacial pad (MP) activity is state-dependent. Schematic diagrams showing the injection protocols for non-selective transduction of preBötC neurons (A) and selective transduction of preBötC→7n neurons (K). Representative trace showing the integrated (∫) and raw MP EMG, heart rate (HR), electrocardiograms (ECG), and ∫ and raw diaphragm (Dia) EMG in the initial phase of reversal of ketamine/medetomidine anesthesia in rats with non-selective (B) and selective (L) preBötC neuron transduction. The period of photoinhibition is highlighted with the blue box. Group data showing mean (solid line) and 95% confidence intervals for (C and M) respiratory frequency - fR, (E and O) Dia amplitude, (G and Q) and HR (bpm) before, during, and after photoinhibition in non-selective (red), selective GtACR2 expressing (blue), and control (black) rats. Histograms showing group data for the effect of photoinhibition of preBötC on respiratory and cardiovascular parameters (D and N) fR, (F and P) Dia amplitude, (H and R) HR, (I and S) apnea duration and number of missed respiratory cycles, and (J and T) MP activity.; P1 and P2 refer to the initial period of photoinhibition. Group data are presented as mean ± 95% CI; unpaired t-test or nonparametric Mann- Whitney test with multiple comparisons using the Bonferroni-Dunn method, *p<0.05. The photoinhibition of preBötC neurons is depicted by blue

Article Snippet: resource Designation Source or reference Identifiers Additional information Antibody Biotin- SP- conjugated donkey polyclonal anti- goat Jackson ImmunoResearch Laboratories, Inc Cat#: 705065147 1:500 Antibody Rabbit polyclonal anti- NK1R Sigma- Aldrich Cat # S8305 1:5000 Antibody Streptavidin- Marina blue Invitrogen- Thermo Fisher Scientific Cat#: S11221 1:200 Commercial assay or kit RNAscope multiplex fluorescence v1 kit ACD Cat #320851 Commercial assay or kit Protease IV ACD Cat #322336 Commercial assay or kit VGAT (SLC32A1) ACD Cat #ADV424541- C2 Commercial assay or kit VGluT2 (Slc17a6) ACD Cat #317011- C3 Commercial assay or kit SST ACD Cat#ADV412181- C3 Commercial assay or kit Reelin ACD Cat ADV1048921- C1 Commercial assay or kit C1 diluent ACD Cat #ADV300041 Chemical compound, drug Meloxicam Lyppard 1 mg/kg, s.c. Chemical compound, drug Isoflurane Rhodia Australia 5% induction, 2–2.5% maintenance Chemical compound, drug Ketamine Lyppard 75 mg/kg, i.p. Chemical compound, drug Medetomidine Pfizer Animal Health 0.5 mg/kg, i.p. Chemical compound, drug Atipamazole Pfizer Animal Health 1 mg/kg, i.p Chemical compound, drug Urethane Sigma- Aldrich 1.4 g/kg, i.v. Chemical compound, drug Buprenorphine Schering- Plough 0.025 mg/kg, s.c. Software, algorithm Spike2 Cambridge Electrical Design Software, algorithm Zen Blue Carl- Zeiss Software, algorithm Zen Black Carl- Zeiss Software, algorithm ImageJ NIH Software, algorithm SigmaPlot v11 Systat Software Inc Software, algorithm Prims v9.0 GraphPad Continued

Techniques: Inhibition, Activity Assay, Injection, Transduction, Expressing, Control, MANN-WHITNEY

Figure 9. Inspiratory-related activity of mystacial pad is interrupted by the selective inhibition of pre-Bötzinger complex (preBötC) to 7n neurons in conscious rats. Schematic diagrams showing the injection protocols for non-selective transduction of preBötC neurons (A) and selective transduction of preBötC→7n neurons (K). Representative trace showing the integrated (∫) and raw mystacial pad (MP) EMG, heart rate (HR), electrocardiograms (ECG), and ∫ and raw diaphragm (Dia) EMG after recovery from ketamine/medetomidine anesthesia in rats with non-selective (B) and selective (L) preBötC neuron transduction. The period of photoinhibition is highlighted with the blue box. Group data showing mean (solid line) and 95% confidence intervals for (C and M) respiratory frequency - fR, (E and O) Dia amplitude, (G and Q) and HR (bpm) before, during, and after photoinhibition in non- selective (red), selective GtACR2 expressing (blue) and control (black) rats. Histograms showing group data for the effect of photoinhibition of preBötC on respiratory and cardiovascular parameters (D and N) fR, (F and P) Dia amplitude, (H and R) HR, (I and S) apnea duration and number of missed respiratory cycles, and (J and T) MP activity; P1 and P2 refer to the initial period of photoinhibition. Group data are presented as mean ± 95% CI; unpaired t-test or nonparametric Mann-Whitney test with multiple comparisons using the Bonferroni-Dunn method, *p<0.05. The photoinhibition of preBötC neurons is depicted by blue shading.

Journal: eLife

Article Title: Selective transduction and photoinhibition of pre-Bötzinger neurons that project to the facial nucleus in rats affect the nasofacial activity

doi: 10.7554/elife.85398

Figure Lengend Snippet: Figure 9. Inspiratory-related activity of mystacial pad is interrupted by the selective inhibition of pre-Bötzinger complex (preBötC) to 7n neurons in conscious rats. Schematic diagrams showing the injection protocols for non-selective transduction of preBötC neurons (A) and selective transduction of preBötC→7n neurons (K). Representative trace showing the integrated (∫) and raw mystacial pad (MP) EMG, heart rate (HR), electrocardiograms (ECG), and ∫ and raw diaphragm (Dia) EMG after recovery from ketamine/medetomidine anesthesia in rats with non-selective (B) and selective (L) preBötC neuron transduction. The period of photoinhibition is highlighted with the blue box. Group data showing mean (solid line) and 95% confidence intervals for (C and M) respiratory frequency - fR, (E and O) Dia amplitude, (G and Q) and HR (bpm) before, during, and after photoinhibition in non- selective (red), selective GtACR2 expressing (blue) and control (black) rats. Histograms showing group data for the effect of photoinhibition of preBötC on respiratory and cardiovascular parameters (D and N) fR, (F and P) Dia amplitude, (H and R) HR, (I and S) apnea duration and number of missed respiratory cycles, and (J and T) MP activity; P1 and P2 refer to the initial period of photoinhibition. Group data are presented as mean ± 95% CI; unpaired t-test or nonparametric Mann-Whitney test with multiple comparisons using the Bonferroni-Dunn method, *p<0.05. The photoinhibition of preBötC neurons is depicted by blue shading.

Article Snippet: resource Designation Source or reference Identifiers Additional information Antibody Biotin- SP- conjugated donkey polyclonal anti- goat Jackson ImmunoResearch Laboratories, Inc Cat#: 705065147 1:500 Antibody Rabbit polyclonal anti- NK1R Sigma- Aldrich Cat # S8305 1:5000 Antibody Streptavidin- Marina blue Invitrogen- Thermo Fisher Scientific Cat#: S11221 1:200 Commercial assay or kit RNAscope multiplex fluorescence v1 kit ACD Cat #320851 Commercial assay or kit Protease IV ACD Cat #322336 Commercial assay or kit VGAT (SLC32A1) ACD Cat #ADV424541- C2 Commercial assay or kit VGluT2 (Slc17a6) ACD Cat #317011- C3 Commercial assay or kit SST ACD Cat#ADV412181- C3 Commercial assay or kit Reelin ACD Cat ADV1048921- C1 Commercial assay or kit C1 diluent ACD Cat #ADV300041 Chemical compound, drug Meloxicam Lyppard 1 mg/kg, s.c. Chemical compound, drug Isoflurane Rhodia Australia 5% induction, 2–2.5% maintenance Chemical compound, drug Ketamine Lyppard 75 mg/kg, i.p. Chemical compound, drug Medetomidine Pfizer Animal Health 0.5 mg/kg, i.p. Chemical compound, drug Atipamazole Pfizer Animal Health 1 mg/kg, i.p Chemical compound, drug Urethane Sigma- Aldrich 1.4 g/kg, i.v. Chemical compound, drug Buprenorphine Schering- Plough 0.025 mg/kg, s.c. Software, algorithm Spike2 Cambridge Electrical Design Software, algorithm Zen Blue Carl- Zeiss Software, algorithm Zen Black Carl- Zeiss Software, algorithm ImageJ NIH Software, algorithm SigmaPlot v11 Systat Software Inc Software, algorithm Prims v9.0 GraphPad Continued

Techniques: Activity Assay, Inhibition, Injection, Transduction, Expressing, Control, MANN-WHITNEY

Figure 4. Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the PI3K/AKT/FOXO1 pathway. Effects of US-CM and GDNF on the phosphorylated FOXO1 (p-FOXO1)/FOXO1 ratio, total FOXO1 expression (A), cytoplasmic p-FOXO1, cytoplasmic FOXO1, and nuclear FOXO1 expression (B). The expression levels of total and nuclear FOXO1 (C), claudin-5, and VE-cadherin (D) in hCMEC/D3 cells transfected with FOXO1 siRNA (siFOXO1). NC: negative control. (E) Effects of FOXO1 overexpression (FOXO1-OE) and GDNF on the expression levels of claudin-5, total FOXO1, and nuclear FOXO1. FOXO1-NC: negative control plasmids. (F) Effects of LY and U0 on GDNF- induced alterations of total p-FOXO1/FOXO1 ratio, cytoplasmic p-FOXO1, cytoplasmic FOXO1, and nuclear FOXO1 expression. (G) Effects of LY on the claudin-5 expression upregulated by siFOXO1. The above data are shown as the mean± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test, Welch’s ANOVA test, or Kruskal– Wallis test.

Journal: eLife

Article Title: Neurons enhance blood–brain barrier function via upregulating claudin-5 and VE-cadherin expression due to glial cell line-derived neurotrophic factor secretion

doi: 10.7554/elife.96161.3

Figure Lengend Snippet: Figure 4. Glial cell line-derived neurotrophic factor (GDNF) induced the claudin-5 expression in hCMEC/D3 cells by activating the PI3K/AKT/FOXO1 pathway. Effects of US-CM and GDNF on the phosphorylated FOXO1 (p-FOXO1)/FOXO1 ratio, total FOXO1 expression (A), cytoplasmic p-FOXO1, cytoplasmic FOXO1, and nuclear FOXO1 expression (B). The expression levels of total and nuclear FOXO1 (C), claudin-5, and VE-cadherin (D) in hCMEC/D3 cells transfected with FOXO1 siRNA (siFOXO1). NC: negative control. (E) Effects of FOXO1 overexpression (FOXO1-OE) and GDNF on the expression levels of claudin-5, total FOXO1, and nuclear FOXO1. FOXO1-NC: negative control plasmids. (F) Effects of LY and U0 on GDNF- induced alterations of total p-FOXO1/FOXO1 ratio, cytoplasmic p-FOXO1, cytoplasmic FOXO1, and nuclear FOXO1 expression. (G) Effects of LY on the claudin-5 expression upregulated by siFOXO1. The above data are shown as the mean± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test, Welch’s ANOVA test, or Kruskal– Wallis test.

Article Snippet: resource Designation Source or reference Identifiers Additional information Antibody p38 (rabbit monoclonal) CST 8690S RRID:AB_10999090 1:250 Antibody p- JNK (rabbit polyclonal) Wanleibio WL01813 RRID:AB_2910628 1:1000 Antibody JNK (rabbit polyclonal) Wanleibio WL01295 RRID:AB_3064853 1:1000 Antibody FOXO1 (rabbit polyclonal) Proteintech 18592 RRID:AB_2934932 1:1000 Antibody p- FOXO1 (rabbit polyclonal) Wanleibio WL03634 RRID: AB_3076326 1:1000 Antibody ETS1 (mouse monoclonal) Santa Cruz sc- 55581 RRID:AB_831289 1:500 Antibody ETS1 (mouse monoclonal) Proteintech 66598 RRID:AB_2881958 1:3000 Cell line (Homo sapiens) hCMEC/D3 cells JENNIO Biological Technology, Guangzhou, China Cat#JNO- H0520 RRID:CVCL_U985 Authenticated (STR profiling) Cell line (H. sapiens) U251 cells Cellcook Biological Technology, Guangzhou, China Cat#CC1701 RRID:CVCL_0021 Authenticated (STR profiling) Cell line (H. sapiens) SH- SY5Y cells Cellcook Biological Technology, Guangzhou, China Cat#CC2101 RRID:CVCL_0019 Authenticated (STR profiling) Software, algorithm GraphPad Prism Version 8.0.2 RRID:SCR_002798 Software, algorithm BioTek Cytation 5 Cell Imaging Multi- Mode Reader BioTek Cytation 5 RRID:SCR_019732 Software, algorithm QuantStudio 3 Real Time PCR System QuantStudio 3 RRID:SCR_018712 Software, algorithm FACS Celesta Flow Cytometer BD Biosciences RRID:SCR_019597 Software, algorithm Flowjo software Version 10.4 RRID:SCR_008520 Commercial assay or kit GDNF- Elisa kit R&D system RRID:SCR_006140 Cat#212- GD Commercial assay or kit bFGF- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H6042 Commercial assay or kit IGF- 1- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H0086 Commercial assay or kit TGF-β- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- 0162 Peptide, recombinant protein GDNF R&D system RRID:SCR_006140 Cat#212- GD Peptide, recombinant protein bFGF MedChemExpress RRID:SCR_025062 Cat#HY- P7331 Peptide, recombinant protein IGF- 1 MedChemExpress RRID:SCR_025062 Cat#HY- P70783 Peptide, recombinant protein TGF-β MedChemExpress RRID:SCR_025062 Cat#HY- P70543 Chemical compound, drug SPP- 86 MedChemExpress RRID:SCR_025062 Cat#HY- 110193 Chemical compound, drug PP2 MedChemExpress RRID:SCR_025062 Cat#HY- 13805 Continued Continued on next page Yang et al. eLife 2024;13:RP96161.

Techniques: Derivative Assay, Expressing, Transfection, Negative Control, Over Expression

Figure 6. The deficiency of brain glial cell line-derived neurotrophic factor (GDNF) in mice increased the permeability of blood–brain barrier (BBB) and reduced claudin-5 and VE-cadherin expression in mice brains. (A) Experimental configuration of AAV-GFP (shNC) or AAV-shGdnf (shGdnf) intracerebroventricular injection. (B) Effects of brain-specific Gdnf silencing on the expression levels of GDNF, claudin-5, and VE-cadherin in the brains. Effects of brain-specific Gdnf silencing on NaF levels in plasma (C), brain (D), and the ratio of brain to plasma (E). Effects of brain-specific Gdnf silencing on FITC-Dex levels in plasma (F), brain (G), and the ratio of brain to plasma (H). The expression ratios of p-AKT/AKT (I), p-ERK/ERK (J), and p-FOXO1/ FOXO1 (K) in the brains of Gdnf silencing mice. (L) The expression level of ETS1 in the brains of Gdnf silencing mice. The above data are shown as the mean ± SEM. Six biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by unpaired t-test, unpaired t-test with Welch’s correction, or Mann–Whitney test.

Journal: eLife

Article Title: Neurons enhance blood–brain barrier function via upregulating claudin-5 and VE-cadherin expression due to glial cell line-derived neurotrophic factor secretion

doi: 10.7554/elife.96161.3

Figure Lengend Snippet: Figure 6. The deficiency of brain glial cell line-derived neurotrophic factor (GDNF) in mice increased the permeability of blood–brain barrier (BBB) and reduced claudin-5 and VE-cadherin expression in mice brains. (A) Experimental configuration of AAV-GFP (shNC) or AAV-shGdnf (shGdnf) intracerebroventricular injection. (B) Effects of brain-specific Gdnf silencing on the expression levels of GDNF, claudin-5, and VE-cadherin in the brains. Effects of brain-specific Gdnf silencing on NaF levels in plasma (C), brain (D), and the ratio of brain to plasma (E). Effects of brain-specific Gdnf silencing on FITC-Dex levels in plasma (F), brain (G), and the ratio of brain to plasma (H). The expression ratios of p-AKT/AKT (I), p-ERK/ERK (J), and p-FOXO1/ FOXO1 (K) in the brains of Gdnf silencing mice. (L) The expression level of ETS1 in the brains of Gdnf silencing mice. The above data are shown as the mean ± SEM. Six biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by unpaired t-test, unpaired t-test with Welch’s correction, or Mann–Whitney test.

Article Snippet: resource Designation Source or reference Identifiers Additional information Antibody p38 (rabbit monoclonal) CST 8690S RRID:AB_10999090 1:250 Antibody p- JNK (rabbit polyclonal) Wanleibio WL01813 RRID:AB_2910628 1:1000 Antibody JNK (rabbit polyclonal) Wanleibio WL01295 RRID:AB_3064853 1:1000 Antibody FOXO1 (rabbit polyclonal) Proteintech 18592 RRID:AB_2934932 1:1000 Antibody p- FOXO1 (rabbit polyclonal) Wanleibio WL03634 RRID: AB_3076326 1:1000 Antibody ETS1 (mouse monoclonal) Santa Cruz sc- 55581 RRID:AB_831289 1:500 Antibody ETS1 (mouse monoclonal) Proteintech 66598 RRID:AB_2881958 1:3000 Cell line (Homo sapiens) hCMEC/D3 cells JENNIO Biological Technology, Guangzhou, China Cat#JNO- H0520 RRID:CVCL_U985 Authenticated (STR profiling) Cell line (H. sapiens) U251 cells Cellcook Biological Technology, Guangzhou, China Cat#CC1701 RRID:CVCL_0021 Authenticated (STR profiling) Cell line (H. sapiens) SH- SY5Y cells Cellcook Biological Technology, Guangzhou, China Cat#CC2101 RRID:CVCL_0019 Authenticated (STR profiling) Software, algorithm GraphPad Prism Version 8.0.2 RRID:SCR_002798 Software, algorithm BioTek Cytation 5 Cell Imaging Multi- Mode Reader BioTek Cytation 5 RRID:SCR_019732 Software, algorithm QuantStudio 3 Real Time PCR System QuantStudio 3 RRID:SCR_018712 Software, algorithm FACS Celesta Flow Cytometer BD Biosciences RRID:SCR_019597 Software, algorithm Flowjo software Version 10.4 RRID:SCR_008520 Commercial assay or kit GDNF- Elisa kit R&D system RRID:SCR_006140 Cat#212- GD Commercial assay or kit bFGF- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H6042 Commercial assay or kit IGF- 1- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H0086 Commercial assay or kit TGF-β- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- 0162 Peptide, recombinant protein GDNF R&D system RRID:SCR_006140 Cat#212- GD Peptide, recombinant protein bFGF MedChemExpress RRID:SCR_025062 Cat#HY- P7331 Peptide, recombinant protein IGF- 1 MedChemExpress RRID:SCR_025062 Cat#HY- P70783 Peptide, recombinant protein TGF-β MedChemExpress RRID:SCR_025062 Cat#HY- P70543 Chemical compound, drug SPP- 86 MedChemExpress RRID:SCR_025062 Cat#HY- 110193 Chemical compound, drug PP2 MedChemExpress RRID:SCR_025062 Cat#HY- 13805 Continued Continued on next page Yang et al. eLife 2024;13:RP96161.

Techniques: Derivative Assay, Permeability, Expressing, Injection, Clinical Proteomics, MANN-WHITNEY

Figure 8. The mechanism of neurons and astrocytes induced the integrity of brain endothelial cells. Neurons but also astrocytes trigger the activation of PI3K/AKT and MAPK/ERK pathways in brain endothelial cells by glial cell line-derived neurotrophic factor (GDNF) secretion, which in turn regulates transcription factors of claudin-5 (FOXO1) and VE-cadherin (ETS1) to promote claudin-5 and VE-cadherin expression and leads to the enhancement of blood–brain barrier (BBB) integrity. Meanwhile, with the increase in barrier integrity, the in vitro BBB model also obtained a stronger in vivo correlation.

Journal: eLife

Article Title: Neurons enhance blood–brain barrier function via upregulating claudin-5 and VE-cadherin expression due to glial cell line-derived neurotrophic factor secretion

doi: 10.7554/elife.96161.3

Figure Lengend Snippet: Figure 8. The mechanism of neurons and astrocytes induced the integrity of brain endothelial cells. Neurons but also astrocytes trigger the activation of PI3K/AKT and MAPK/ERK pathways in brain endothelial cells by glial cell line-derived neurotrophic factor (GDNF) secretion, which in turn regulates transcription factors of claudin-5 (FOXO1) and VE-cadherin (ETS1) to promote claudin-5 and VE-cadherin expression and leads to the enhancement of blood–brain barrier (BBB) integrity. Meanwhile, with the increase in barrier integrity, the in vitro BBB model also obtained a stronger in vivo correlation.

Article Snippet: resource Designation Source or reference Identifiers Additional information Antibody p38 (rabbit monoclonal) CST 8690S RRID:AB_10999090 1:250 Antibody p- JNK (rabbit polyclonal) Wanleibio WL01813 RRID:AB_2910628 1:1000 Antibody JNK (rabbit polyclonal) Wanleibio WL01295 RRID:AB_3064853 1:1000 Antibody FOXO1 (rabbit polyclonal) Proteintech 18592 RRID:AB_2934932 1:1000 Antibody p- FOXO1 (rabbit polyclonal) Wanleibio WL03634 RRID: AB_3076326 1:1000 Antibody ETS1 (mouse monoclonal) Santa Cruz sc- 55581 RRID:AB_831289 1:500 Antibody ETS1 (mouse monoclonal) Proteintech 66598 RRID:AB_2881958 1:3000 Cell line (Homo sapiens) hCMEC/D3 cells JENNIO Biological Technology, Guangzhou, China Cat#JNO- H0520 RRID:CVCL_U985 Authenticated (STR profiling) Cell line (H. sapiens) U251 cells Cellcook Biological Technology, Guangzhou, China Cat#CC1701 RRID:CVCL_0021 Authenticated (STR profiling) Cell line (H. sapiens) SH- SY5Y cells Cellcook Biological Technology, Guangzhou, China Cat#CC2101 RRID:CVCL_0019 Authenticated (STR profiling) Software, algorithm GraphPad Prism Version 8.0.2 RRID:SCR_002798 Software, algorithm BioTek Cytation 5 Cell Imaging Multi- Mode Reader BioTek Cytation 5 RRID:SCR_019732 Software, algorithm QuantStudio 3 Real Time PCR System QuantStudio 3 RRID:SCR_018712 Software, algorithm FACS Celesta Flow Cytometer BD Biosciences RRID:SCR_019597 Software, algorithm Flowjo software Version 10.4 RRID:SCR_008520 Commercial assay or kit GDNF- Elisa kit R&D system RRID:SCR_006140 Cat#212- GD Commercial assay or kit bFGF- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H6042 Commercial assay or kit IGF- 1- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H0086 Commercial assay or kit TGF-β- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- 0162 Peptide, recombinant protein GDNF R&D system RRID:SCR_006140 Cat#212- GD Peptide, recombinant protein bFGF MedChemExpress RRID:SCR_025062 Cat#HY- P7331 Peptide, recombinant protein IGF- 1 MedChemExpress RRID:SCR_025062 Cat#HY- P70783 Peptide, recombinant protein TGF-β MedChemExpress RRID:SCR_025062 Cat#HY- P70543 Chemical compound, drug SPP- 86 MedChemExpress RRID:SCR_025062 Cat#HY- 110193 Chemical compound, drug PP2 MedChemExpress RRID:SCR_025062 Cat#HY- 13805 Continued Continued on next page Yang et al. eLife 2024;13:RP96161.

Techniques: Activation Assay, Derivative Assay, Expressing, In Vitro, In Vivo

Figure 5. Glial cell line-derived neurotrophic factor (GDNF)-induced VE-cadherin expression in hCMEC/D3 cells by activating the PI3K/AKT/ETS1 and MAPK/ERK/ETS1 pathways. Effects of US-CM and GDNF on total (A) and nuclear (B) ETS1 expression. Effects of LY and U0 on 200 pg/ml GDNF-induced total (C) and nuclear (D) ETS1 expression. Expression levels of total (E) and the nuclear ETS1 (F) in hCMEC/D3 cells after knocking down ETS1 with siRNA (siETS1). (G) Effects of GDNF and siETS1 on the expression of VE-cadherin and claudin-5. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test.

Journal: eLife

Article Title: Neurons enhance blood–brain barrier function via upregulating claudin-5 and VE-cadherin expression due to glial cell line-derived neurotrophic factor secretion

doi: 10.7554/elife.96161.3

Figure Lengend Snippet: Figure 5. Glial cell line-derived neurotrophic factor (GDNF)-induced VE-cadherin expression in hCMEC/D3 cells by activating the PI3K/AKT/ETS1 and MAPK/ERK/ETS1 pathways. Effects of US-CM and GDNF on total (A) and nuclear (B) ETS1 expression. Effects of LY and U0 on 200 pg/ml GDNF-induced total (C) and nuclear (D) ETS1 expression. Expression levels of total (E) and the nuclear ETS1 (F) in hCMEC/D3 cells after knocking down ETS1 with siRNA (siETS1). (G) Effects of GDNF and siETS1 on the expression of VE-cadherin and claudin-5. The above data are shown as the mean ± SEM. Four biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by one-way ANOVA test followed by Fisher’s LSD test.

Article Snippet: resource Designation Source or reference Identifiers Additional information Antibody p38 (rabbit monoclonal) CST 8690S RRID:AB_10999090 1:250 Antibody p- JNK (rabbit polyclonal) Wanleibio WL01813 RRID:AB_2910628 1:1000 Antibody JNK (rabbit polyclonal) Wanleibio WL01295 RRID:AB_3064853 1:1000 Antibody FOXO1 (rabbit polyclonal) Proteintech 18592 RRID:AB_2934932 1:1000 Antibody p- FOXO1 (rabbit polyclonal) Wanleibio WL03634 RRID: AB_3076326 1:1000 Antibody ETS1 (mouse monoclonal) Santa Cruz sc- 55581 RRID:AB_831289 1:500 Antibody ETS1 (mouse monoclonal) Proteintech 66598 RRID:AB_2881958 1:3000 Cell line (Homo sapiens) hCMEC/D3 cells JENNIO Biological Technology, Guangzhou, China Cat#JNO- H0520 RRID:CVCL_U985 Authenticated (STR profiling) Cell line (H. sapiens) U251 cells Cellcook Biological Technology, Guangzhou, China Cat#CC1701 RRID:CVCL_0021 Authenticated (STR profiling) Cell line (H. sapiens) SH- SY5Y cells Cellcook Biological Technology, Guangzhou, China Cat#CC2101 RRID:CVCL_0019 Authenticated (STR profiling) Software, algorithm GraphPad Prism Version 8.0.2 RRID:SCR_002798 Software, algorithm BioTek Cytation 5 Cell Imaging Multi- Mode Reader BioTek Cytation 5 RRID:SCR_019732 Software, algorithm QuantStudio 3 Real Time PCR System QuantStudio 3 RRID:SCR_018712 Software, algorithm FACS Celesta Flow Cytometer BD Biosciences RRID:SCR_019597 Software, algorithm Flowjo software Version 10.4 RRID:SCR_008520 Commercial assay or kit GDNF- Elisa kit R&D system RRID:SCR_006140 Cat#212- GD Commercial assay or kit bFGF- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H6042 Commercial assay or kit IGF- 1- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H0086 Commercial assay or kit TGF-β- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- 0162 Peptide, recombinant protein GDNF R&D system RRID:SCR_006140 Cat#212- GD Peptide, recombinant protein bFGF MedChemExpress RRID:SCR_025062 Cat#HY- P7331 Peptide, recombinant protein IGF- 1 MedChemExpress RRID:SCR_025062 Cat#HY- P70783 Peptide, recombinant protein TGF-β MedChemExpress RRID:SCR_025062 Cat#HY- P70543 Chemical compound, drug SPP- 86 MedChemExpress RRID:SCR_025062 Cat#HY- 110193 Chemical compound, drug PP2 MedChemExpress RRID:SCR_025062 Cat#HY- 13805 Continued Continued on next page Yang et al. eLife 2024;13:RP96161.

Techniques: Derivative Assay, Expressing

Figure 6. The deficiency of brain glial cell line-derived neurotrophic factor (GDNF) in mice increased the permeability of blood–brain barrier (BBB) and reduced claudin-5 and VE-cadherin expression in mice brains. (A) Experimental configuration of AAV-GFP (shNC) or AAV-shGdnf (shGdnf) intracerebroventricular injection. (B) Effects of brain-specific Gdnf silencing on the expression levels of GDNF, claudin-5, and VE-cadherin in the brains. Effects of brain-specific Gdnf silencing on NaF levels in plasma (C), brain (D), and the ratio of brain to plasma (E). Effects of brain-specific Gdnf silencing on FITC-Dex levels in plasma (F), brain (G), and the ratio of brain to plasma (H). The expression ratios of p-AKT/AKT (I), p-ERK/ERK (J), and p-FOXO1/ FOXO1 (K) in the brains of Gdnf silencing mice. (L) The expression level of ETS1 in the brains of Gdnf silencing mice. The above data are shown as the mean ± SEM. Six biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by unpaired t-test, unpaired t-test with Welch’s correction, or Mann–Whitney test.

Journal: eLife

Article Title: Neurons enhance blood–brain barrier function via upregulating claudin-5 and VE-cadherin expression due to glial cell line-derived neurotrophic factor secretion

doi: 10.7554/elife.96161.3

Figure Lengend Snippet: Figure 6. The deficiency of brain glial cell line-derived neurotrophic factor (GDNF) in mice increased the permeability of blood–brain barrier (BBB) and reduced claudin-5 and VE-cadherin expression in mice brains. (A) Experimental configuration of AAV-GFP (shNC) or AAV-shGdnf (shGdnf) intracerebroventricular injection. (B) Effects of brain-specific Gdnf silencing on the expression levels of GDNF, claudin-5, and VE-cadherin in the brains. Effects of brain-specific Gdnf silencing on NaF levels in plasma (C), brain (D), and the ratio of brain to plasma (E). Effects of brain-specific Gdnf silencing on FITC-Dex levels in plasma (F), brain (G), and the ratio of brain to plasma (H). The expression ratios of p-AKT/AKT (I), p-ERK/ERK (J), and p-FOXO1/ FOXO1 (K) in the brains of Gdnf silencing mice. (L) The expression level of ETS1 in the brains of Gdnf silencing mice. The above data are shown as the mean ± SEM. Six biological replicates per group. One technical replicate for each biological replicate. *p < 0.05; **p < 0.01 by unpaired t-test, unpaired t-test with Welch’s correction, or Mann–Whitney test.

Article Snippet: resource Designation Source or reference Identifiers Additional information Antibody p38 (rabbit monoclonal) CST 8690S RRID:AB_10999090 1:250 Antibody p- JNK (rabbit polyclonal) Wanleibio WL01813 RRID:AB_2910628 1:1000 Antibody JNK (rabbit polyclonal) Wanleibio WL01295 RRID:AB_3064853 1:1000 Antibody FOXO1 (rabbit polyclonal) Proteintech 18592 RRID:AB_2934932 1:1000 Antibody p- FOXO1 (rabbit polyclonal) Wanleibio WL03634 RRID: AB_3076326 1:1000 Antibody ETS1 (mouse monoclonal) Santa Cruz sc- 55581 RRID:AB_831289 1:500 Antibody ETS1 (mouse monoclonal) Proteintech 66598 RRID:AB_2881958 1:3000 Cell line (Homo sapiens) hCMEC/D3 cells JENNIO Biological Technology, Guangzhou, China Cat#JNO- H0520 RRID:CVCL_U985 Authenticated (STR profiling) Cell line (H. sapiens) U251 cells Cellcook Biological Technology, Guangzhou, China Cat#CC1701 RRID:CVCL_0021 Authenticated (STR profiling) Cell line (H. sapiens) SH- SY5Y cells Cellcook Biological Technology, Guangzhou, China Cat#CC2101 RRID:CVCL_0019 Authenticated (STR profiling) Software, algorithm GraphPad Prism Version 8.0.2 RRID:SCR_002798 Software, algorithm BioTek Cytation 5 Cell Imaging Multi- Mode Reader BioTek Cytation 5 RRID:SCR_019732 Software, algorithm QuantStudio 3 Real Time PCR System QuantStudio 3 RRID:SCR_018712 Software, algorithm FACS Celesta Flow Cytometer BD Biosciences RRID:SCR_019597 Software, algorithm Flowjo software Version 10.4 RRID:SCR_008520 Commercial assay or kit GDNF- Elisa kit R&D system RRID:SCR_006140 Cat#212- GD Commercial assay or kit bFGF- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H6042 Commercial assay or kit IGF- 1- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H0086 Commercial assay or kit TGF-β- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- 0162 Peptide, recombinant protein GDNF R&D system RRID:SCR_006140 Cat#212- GD Peptide, recombinant protein bFGF MedChemExpress RRID:SCR_025062 Cat#HY- P7331 Peptide, recombinant protein IGF- 1 MedChemExpress RRID:SCR_025062 Cat#HY- P70783 Peptide, recombinant protein TGF-β MedChemExpress RRID:SCR_025062 Cat#HY- P70543 Chemical compound, drug SPP- 86 MedChemExpress RRID:SCR_025062 Cat#HY- 110193 Chemical compound, drug PP2 MedChemExpress RRID:SCR_025062 Cat#HY- 13805 Continued Continued on next page Yang et al. eLife 2024;13:RP96161.

Techniques: Derivative Assay, Permeability, Expressing, Injection, Clinical Proteomics, MANN-WHITNEY

Figure 8. The mechanism of neurons and astrocytes induced the integrity of brain endothelial cells. Neurons but also astrocytes trigger the activation of PI3K/AKT and MAPK/ERK pathways in brain endothelial cells by glial cell line-derived neurotrophic factor (GDNF) secretion, which in turn regulates transcription factors of claudin-5 (FOXO1) and VE-cadherin (ETS1) to promote claudin-5 and VE-cadherin expression and leads to the enhancement of blood–brain barrier (BBB) integrity. Meanwhile, with the increase in barrier integrity, the in vitro BBB model also obtained a stronger in vivo correlation.

Journal: eLife

Article Title: Neurons enhance blood–brain barrier function via upregulating claudin-5 and VE-cadherin expression due to glial cell line-derived neurotrophic factor secretion

doi: 10.7554/elife.96161.3

Figure Lengend Snippet: Figure 8. The mechanism of neurons and astrocytes induced the integrity of brain endothelial cells. Neurons but also astrocytes trigger the activation of PI3K/AKT and MAPK/ERK pathways in brain endothelial cells by glial cell line-derived neurotrophic factor (GDNF) secretion, which in turn regulates transcription factors of claudin-5 (FOXO1) and VE-cadherin (ETS1) to promote claudin-5 and VE-cadherin expression and leads to the enhancement of blood–brain barrier (BBB) integrity. Meanwhile, with the increase in barrier integrity, the in vitro BBB model also obtained a stronger in vivo correlation.

Article Snippet: resource Designation Source or reference Identifiers Additional information Antibody p38 (rabbit monoclonal) CST 8690S RRID:AB_10999090 1:250 Antibody p- JNK (rabbit polyclonal) Wanleibio WL01813 RRID:AB_2910628 1:1000 Antibody JNK (rabbit polyclonal) Wanleibio WL01295 RRID:AB_3064853 1:1000 Antibody FOXO1 (rabbit polyclonal) Proteintech 18592 RRID:AB_2934932 1:1000 Antibody p- FOXO1 (rabbit polyclonal) Wanleibio WL03634 RRID: AB_3076326 1:1000 Antibody ETS1 (mouse monoclonal) Santa Cruz sc- 55581 RRID:AB_831289 1:500 Antibody ETS1 (mouse monoclonal) Proteintech 66598 RRID:AB_2881958 1:3000 Cell line (Homo sapiens) hCMEC/D3 cells JENNIO Biological Technology, Guangzhou, China Cat#JNO- H0520 RRID:CVCL_U985 Authenticated (STR profiling) Cell line (H. sapiens) U251 cells Cellcook Biological Technology, Guangzhou, China Cat#CC1701 RRID:CVCL_0021 Authenticated (STR profiling) Cell line (H. sapiens) SH- SY5Y cells Cellcook Biological Technology, Guangzhou, China Cat#CC2101 RRID:CVCL_0019 Authenticated (STR profiling) Software, algorithm GraphPad Prism Version 8.0.2 RRID:SCR_002798 Software, algorithm BioTek Cytation 5 Cell Imaging Multi- Mode Reader BioTek Cytation 5 RRID:SCR_019732 Software, algorithm QuantStudio 3 Real Time PCR System QuantStudio 3 RRID:SCR_018712 Software, algorithm FACS Celesta Flow Cytometer BD Biosciences RRID:SCR_019597 Software, algorithm Flowjo software Version 10.4 RRID:SCR_008520 Commercial assay or kit GDNF- Elisa kit R&D system RRID:SCR_006140 Cat#212- GD Commercial assay or kit bFGF- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H6042 Commercial assay or kit IGF- 1- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- H0086 Commercial assay or kit TGF-β- Elisa kit Elabscience RRID:SCR_025982 Cat#E- EL- 0162 Peptide, recombinant protein GDNF R&D system RRID:SCR_006140 Cat#212- GD Peptide, recombinant protein bFGF MedChemExpress RRID:SCR_025062 Cat#HY- P7331 Peptide, recombinant protein IGF- 1 MedChemExpress RRID:SCR_025062 Cat#HY- P70783 Peptide, recombinant protein TGF-β MedChemExpress RRID:SCR_025062 Cat#HY- P70543 Chemical compound, drug SPP- 86 MedChemExpress RRID:SCR_025062 Cat#HY- 110193 Chemical compound, drug PP2 MedChemExpress RRID:SCR_025062 Cat#HY- 13805 Continued Continued on next page Yang et al. eLife 2024;13:RP96161.

Techniques: Activation Assay, Derivative Assay, Expressing, In Vitro, In Vivo