Journal: Nanomaterials

Article Title: You Don’t Learn That in School: An Updated Practical Guide to Carbon Quantum Dots

doi: 10.3390/nano11030611

Figure Lengend Snippet: Summarized examples of carbon quantum dots used for biomedical applications.

Article Snippet: Skin permeability , methicillin-sensitive Staphylococcus aureus (ATCC 6538) methicillin-resistant Staphylococcus aureus (ATCC 4300) Escherichia coli (ATCC 25922) Staphylococcus epidermidis (ATCC 49134) Bacillus cereus (ATCC 14579) , n/a , n/a , Hydrothermal , polyethyleneimine (PEI) citric acid (CA) , 250 °C, 4 h , N-doping , Dialysis (≥12,000 Da, 24 h) , 70–10 , PEI:CA ratio (1:0,5)–31 (1:1)–53 (1:2)–7.6 , , [ ] .

Techniques: Purification, In Vivo, In Vitro, Centrifugation, Filtration, Membrane, Bacteria, Imaging, Column Chromatography, Conjugation Assay, Reflux, Polymer, Virus, Modification, Concentration Assay, Biomarker Discovery, Permeability, Isolation

Journal: Nanomaterials

Article Title: You Don’t Learn That in School: An Updated Practical Guide to Carbon Quantum Dots

doi: 10.3390/nano11030611

Figure Lengend Snippet: Summarized examples of carbon quantum dots used as antimicrobials and antivirals.

Article Snippet: Skin permeability , methicillin-sensitive Staphylococcus aureus (ATCC 6538) methicillin-resistant Staphylococcus aureus (ATCC 4300) Escherichia coli (ATCC 25922) Staphylococcus epidermidis (ATCC 49134) Bacillus cereus (ATCC 14579) , n/a , n/a , Hydrothermal , polyethyleneimine (PEI) citric acid (CA) , 250 °C, 4 h , N-doping , Dialysis (≥12,000 Da, 24 h) , 70–10 , PEI:CA ratio (1:0,5)–31 (1:1)–53 (1:2)–7.6 , , [ ] .

Techniques: Bacteria, Centrifugation, Filtration, Membrane, Virus, Reflux, Polymer, Encapsulation

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 1. Meningitic E. coli induction of IL-22 contributes to the degradation of TJPs in hBMEC. (A) ELISA analysis of IL-22 in serum and brain lysates from challenged mice (n = 3). Data were collected and presented as mean ± standard deviation (SD) from three replicates at each time point. *p < 0.05, **p < 0.01, ***p < 0.001. (B) TEER value changes in hBMEC when treated with multiple doses of IL-22 (0, 1, 5, 10, 20, and 100 ng/mL) monitored by the ECIS system. Data were collected and presented as mean ± SD from three replicated wells at each time point. (C) Western blot analysis of ZO-1, Occludin, and Claudin-5 in hBMEC in response to multiple doses of IL-22 (0, 1, 5, 10, 20, and 100 ng/mL). β-actin was used as the loading control. Densitometry was performed to analyze differences among the samples. *p < 0.05, **p < 0.01, ***p < 0.001. (D) Immunofluorescence analysis determining the distribution and expression of ZO-1, Occludin, and Claudin-5 in hBMEC after 10 ng/mL IL-22 treatment. ZO-1, Occludin, and Claudin-5 were stained in red. The cell nucleus was stained in blue with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar, 10 μm.

Article Snippet: The cells were then incubated with the primary ZO-1 (Abcam, ab216880, 1:200), Occludin (Proteintech, 27260-1- AP, 1:400), or Claudin-5 (Abclonal, A10207, 1:100) antibodies for 12 h at 4 °C, and then incubated with goat antirabbit IgG Cy3 antibody (Proteintech, SA00009-2, 1:200) for 1 h. Cells were counterstained with DAPI (Beyotime, Shanghai, China) to visualize nuclear morphology and photographed using a BX41 fluorescence microscope (Olympus, Tokyo, Japan).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Western Blot, Control, Immunofluorescence, Expressing, Staining

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 2. IL-22 promotes BBB permeability enhancement in vivo. (A) Effects of the recombinant mouse IL-22 (10 ng/mouse) on the permeability of the mice brain evaluated using the Evans blue approach (n = 5). (B) Western blot analysis of ZO-1, Occludin, and Claudin-5 in the brains of IL- 22-treated (10 ng/mouse) mice (n = 3). β-actin was used as the loading control. Densitometry was performed to analyze differences among the samples. **p < 0.01, ***p < 0.001. (C) Brain samples of both mice treated with and without IL-22 (10 ng/mouse) for 24 h were analyzed for vascular endothelium integrity by immunofluorescence. The figures represent the results of three mouse brains (three images per mouse). ZO-1, Occludin, and Claudin-5 were selected as the markers reflecting the vascular endothelium integrity. CD31 was specifically applied for labeling the microvessels in green. The cell nucleus was stained in blue with DAPI. Scale bar indicates 50 μm.

Article Snippet: The cells were then incubated with the primary ZO-1 (Abcam, ab216880, 1:200), Occludin (Proteintech, 27260-1- AP, 1:400), or Claudin-5 (Abclonal, A10207, 1:100) antibodies for 12 h at 4 °C, and then incubated with goat antirabbit IgG Cy3 antibody (Proteintech, SA00009-2, 1:200) for 1 h. Cells were counterstained with DAPI (Beyotime, Shanghai, China) to visualize nuclear morphology and photographed using a BX41 fluorescence microscope (Olympus, Tokyo, Japan).

Techniques: Permeability, In Vivo, Recombinant, Western Blot, Control, Immunofluorescence, Labeling, Staining

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 3. IL-22 knockout in mice suppresses meningitic E. coli-induced BBB disruption. (A) Evans blue assay was used to evaluate the permeability of WT and IL-22−/−mice with or without meningitic E. coli infection (n = 5). (B) Brain lysates from the challenged WT and IL-22−/−mice were assessed using Western blot for the expression of TJPs (n = 3). β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. *p < 0.05, **p < 0.01, ***p < 0.001. (C) Immunofluorescence analysis of vascular endothelium integrity in infected WT and IL-22−/−mice. The figures represent the results of three mouse brains (three images per mouse). ZO-1, Occludin, and Claudin-5 were stained in red. CD31 was specifically applied for labeling the microvessels in green. The cell nucleus was stained in blue with DAPI. Scale bar indicates 50 μm.

Article Snippet: The cells were then incubated with the primary ZO-1 (Abcam, ab216880, 1:200), Occludin (Proteintech, 27260-1- AP, 1:400), or Claudin-5 (Abclonal, A10207, 1:100) antibodies for 12 h at 4 °C, and then incubated with goat antirabbit IgG Cy3 antibody (Proteintech, SA00009-2, 1:200) for 1 h. Cells were counterstained with DAPI (Beyotime, Shanghai, China) to visualize nuclear morphology and photographed using a BX41 fluorescence microscope (Olympus, Tokyo, Japan).

Techniques: Knock-Out, Disruption, Evans Blue Assay, Permeability, Infection, Western Blot, Expressing, Control, Immunofluorescence, Staining, Labeling

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 4. IL-22 negatively regulates BBB integrity by activating STAT3. (A) Western blot analysis of STAT3 phosphorylation in hBMEC in response to multiple doses of IL-22 (0, 1, 5, 10, 20, and 100 ng/mL). β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. ***p < 0.001. (B) ECIS system monitoring TEER changes of the IL-22-treated hBMEC with or without STAT3 specific inhibitor S3I-201 administration. Data were collected and presented as mean ± SD from three independently replicated wells at each time point. (C) Effects of the S3I-201 (50 μM) treatment on IL-22-induced (10 ng/mL) downregulation of ZO-1, Occludin, and Claudin-5 in hBMEC. β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. **p < 0.01, ***p < 0.001. (D) Distribution of TJPs in IL-22-treated hBMEC with or without S3I-201 (50 μM) administration, as determined by immunofluorescence analysis. ZO-1, Occludin, and Claudin-5 were stained in red. The cell nucleus was stained in blue with DAPI. Scale bar, 10 μm.

Article Snippet: The cells were then incubated with the primary ZO-1 (Abcam, ab216880, 1:200), Occludin (Proteintech, 27260-1- AP, 1:400), or Claudin-5 (Abclonal, A10207, 1:100) antibodies for 12 h at 4 °C, and then incubated with goat antirabbit IgG Cy3 antibody (Proteintech, SA00009-2, 1:200) for 1 h. Cells were counterstained with DAPI (Beyotime, Shanghai, China) to visualize nuclear morphology and photographed using a BX41 fluorescence microscope (Olympus, Tokyo, Japan).

Techniques: Western Blot, Phospho-proteomics, Control, Immunofluorescence, Staining

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 6. BBB damage caused by IL-22 depends on the upregulation of VEGFA. (A) VEGFA deleted in hBMEC through CRISPR/Cas9, and VEGFA knockout in hBMEC was identified by PCR amplification and Western blot analysis. (B) ECIS assay analyzing the barrier resistance of the WT and VEGFA knockout in hBMEC with the addition of IL-22 (10 ng/mL). (C) Western blot analysis of ZO-1, Occludin, and Claudin-5 expression in the WT and VEGFA knockout in hBMEC in response to IL-22 (10 ng/mL). β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. *p < 0.05, ***p < 0.001. (D) Immunofluorescence analysis determining the distribution and the expression of TJPs in the WT and VEGFA knockout in hBMEC after 10 ng/mL IL-22 treatment. ZO-1, Occludin, and Claudin-5 were stained in red. The cell nucleus was stained in blue with DAPI. Scale bar, 10 μm.

Article Snippet: The cells were then incubated with the primary ZO-1 (Abcam, ab216880, 1:200), Occludin (Proteintech, 27260-1- AP, 1:400), or Claudin-5 (Abclonal, A10207, 1:100) antibodies for 12 h at 4 °C, and then incubated with goat antirabbit IgG Cy3 antibody (Proteintech, SA00009-2, 1:200) for 1 h. Cells were counterstained with DAPI (Beyotime, Shanghai, China) to visualize nuclear morphology and photographed using a BX41 fluorescence microscope (Olympus, Tokyo, Japan).

Techniques: CRISPR, Knock-Out, Amplification, Western Blot, Expressing, Control, Immunofluorescence, Staining

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 1. Meningitic E. coli induction of IL-22 contributes to the degradation of TJPs in hBMEC. (A) ELISA analysis of IL-22 in serum and brain lysates from challenged mice (n = 3). Data were collected and presented as mean ± standard deviation (SD) from three replicates at each time point. *p < 0.05, **p < 0.01, ***p < 0.001. (B) TEER value changes in hBMEC when treated with multiple doses of IL-22 (0, 1, 5, 10, 20, and 100 ng/mL) monitored by the ECIS system. Data were collected and presented as mean ± SD from three replicated wells at each time point. (C) Western blot analysis of ZO-1, Occludin, and Claudin-5 in hBMEC in response to multiple doses of IL-22 (0, 1, 5, 10, 20, and 100 ng/mL). β-actin was used as the loading control. Densitometry was performed to analyze differences among the samples. *p < 0.05, **p < 0.01, ***p < 0.001. (D) Immunofluorescence analysis determining the distribution and expression of ZO-1, Occludin, and Claudin-5 in hBMEC after 10 ng/mL IL-22 treatment. ZO-1, Occludin, and Claudin-5 were stained in red. The cell nucleus was stained in blue with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar, 10 μm.

Article Snippet: ZO-1 (ab216880, 1:1000) and Claudin-5 (ab131259, 1:2000) antibodies were obtained from Abcam (Cambridge, MA). β-actin (66009-1-Ig, 1:50000), Occludin (13409-1-AP, 1:2000), VEGFA (19003-1-AP, 1:1000), and STAT3 (60199-1-Ig, 1:5000) antibodies were obtained from Proteintech (Chicago, IL). p-STAT3 (9145S, 1:2000) was obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Western Blot, Control, Immunofluorescence, Expressing, Staining

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 2. IL-22 promotes BBB permeability enhancement in vivo. (A) Effects of the recombinant mouse IL-22 (10 ng/mouse) on the permeability of the mice brain evaluated using the Evans blue approach (n = 5). (B) Western blot analysis of ZO-1, Occludin, and Claudin-5 in the brains of IL- 22-treated (10 ng/mouse) mice (n = 3). β-actin was used as the loading control. Densitometry was performed to analyze differences among the samples. **p < 0.01, ***p < 0.001. (C) Brain samples of both mice treated with and without IL-22 (10 ng/mouse) for 24 h were analyzed for vascular endothelium integrity by immunofluorescence. The figures represent the results of three mouse brains (three images per mouse). ZO-1, Occludin, and Claudin-5 were selected as the markers reflecting the vascular endothelium integrity. CD31 was specifically applied for labeling the microvessels in green. The cell nucleus was stained in blue with DAPI. Scale bar indicates 50 μm.

Article Snippet: ZO-1 (ab216880, 1:1000) and Claudin-5 (ab131259, 1:2000) antibodies were obtained from Abcam (Cambridge, MA). β-actin (66009-1-Ig, 1:50000), Occludin (13409-1-AP, 1:2000), VEGFA (19003-1-AP, 1:1000), and STAT3 (60199-1-Ig, 1:5000) antibodies were obtained from Proteintech (Chicago, IL). p-STAT3 (9145S, 1:2000) was obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Permeability, In Vivo, Recombinant, Western Blot, Control, Immunofluorescence, Labeling, Staining

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 3. IL-22 knockout in mice suppresses meningitic E. coli-induced BBB disruption. (A) Evans blue assay was used to evaluate the permeability of WT and IL-22−/−mice with or without meningitic E. coli infection (n = 5). (B) Brain lysates from the challenged WT and IL-22−/−mice were assessed using Western blot for the expression of TJPs (n = 3). β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. *p < 0.05, **p < 0.01, ***p < 0.001. (C) Immunofluorescence analysis of vascular endothelium integrity in infected WT and IL-22−/−mice. The figures represent the results of three mouse brains (three images per mouse). ZO-1, Occludin, and Claudin-5 were stained in red. CD31 was specifically applied for labeling the microvessels in green. The cell nucleus was stained in blue with DAPI. Scale bar indicates 50 μm.

Article Snippet: ZO-1 (ab216880, 1:1000) and Claudin-5 (ab131259, 1:2000) antibodies were obtained from Abcam (Cambridge, MA). β-actin (66009-1-Ig, 1:50000), Occludin (13409-1-AP, 1:2000), VEGFA (19003-1-AP, 1:1000), and STAT3 (60199-1-Ig, 1:5000) antibodies were obtained from Proteintech (Chicago, IL). p-STAT3 (9145S, 1:2000) was obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Knock-Out, Disruption, Evans Blue Assay, Permeability, Infection, Western Blot, Expressing, Control, Immunofluorescence, Staining, Labeling

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 4. IL-22 negatively regulates BBB integrity by activating STAT3. (A) Western blot analysis of STAT3 phosphorylation in hBMEC in response to multiple doses of IL-22 (0, 1, 5, 10, 20, and 100 ng/mL). β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. ***p < 0.001. (B) ECIS system monitoring TEER changes of the IL-22-treated hBMEC with or without STAT3 specific inhibitor S3I-201 administration. Data were collected and presented as mean ± SD from three independently replicated wells at each time point. (C) Effects of the S3I-201 (50 μM) treatment on IL-22-induced (10 ng/mL) downregulation of ZO-1, Occludin, and Claudin-5 in hBMEC. β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. **p < 0.01, ***p < 0.001. (D) Distribution of TJPs in IL-22-treated hBMEC with or without S3I-201 (50 μM) administration, as determined by immunofluorescence analysis. ZO-1, Occludin, and Claudin-5 were stained in red. The cell nucleus was stained in blue with DAPI. Scale bar, 10 μm.

Article Snippet: ZO-1 (ab216880, 1:1000) and Claudin-5 (ab131259, 1:2000) antibodies were obtained from Abcam (Cambridge, MA). β-actin (66009-1-Ig, 1:50000), Occludin (13409-1-AP, 1:2000), VEGFA (19003-1-AP, 1:1000), and STAT3 (60199-1-Ig, 1:5000) antibodies were obtained from Proteintech (Chicago, IL). p-STAT3 (9145S, 1:2000) was obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Western Blot, Phospho-proteomics, Control, Immunofluorescence, Staining

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 5. Effects of the IL-22/STAT3 axis on VEGFA expression. (A) Western blot analysis of VEGFA expression in hBMEC in response to multiple doses of IL-22 (0, 1, 2, 5, and 10 ng/mL). β-actin was used as the loading control. Densitometry was performed to analyze differences among the samples. **p < 0.01. (B) ELISA analysis of VEGFA in serum and brain lysates from challenged WT and IL-22−/−mice. Data were collected and presented as mean ± SD *p < 0.05, ***p < 0.001. (C) ChIP-PCR validation of the STAT3 binding to the VEGFA promoter in hBMEC treated with 10 ng/mL IL-22. The nuclear extracts were immunoprecipitated with anti-p-STAT3 antibody or normal IgG and a ChIP assay was performed. The PCR primers were amplified in the −913 to −783 region of the VEGF promoter. Input refers to the same dose of nuclear extract administered prior to immunoprecipitation. (D) Western blot analysis of the effect of S3I-201 (50 μM) treatment on the IL-22-increased (10 ng/mL) VEGFA levels in hBMEC. β-actin was used as the loading control. Densitometry was performed to analyze differences among the samples. **p < 0.01.

Article Snippet: ZO-1 (ab216880, 1:1000) and Claudin-5 (ab131259, 1:2000) antibodies were obtained from Abcam (Cambridge, MA). β-actin (66009-1-Ig, 1:50000), Occludin (13409-1-AP, 1:2000), VEGFA (19003-1-AP, 1:1000), and STAT3 (60199-1-Ig, 1:5000) antibodies were obtained from Proteintech (Chicago, IL). p-STAT3 (9145S, 1:2000) was obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Binding Assay, Immunoprecipitation, Amplification

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 6. BBB damage caused by IL-22 depends on the upregulation of VEGFA. (A) VEGFA deleted in hBMEC through CRISPR/Cas9, and VEGFA knockout in hBMEC was identified by PCR amplification and Western blot analysis. (B) ECIS assay analyzing the barrier resistance of the WT and VEGFA knockout in hBMEC with the addition of IL-22 (10 ng/mL). (C) Western blot analysis of ZO-1, Occludin, and Claudin-5 expression in the WT and VEGFA knockout in hBMEC in response to IL-22 (10 ng/mL). β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. *p < 0.05, ***p < 0.001. (D) Immunofluorescence analysis determining the distribution and the expression of TJPs in the WT and VEGFA knockout in hBMEC after 10 ng/mL IL-22 treatment. ZO-1, Occludin, and Claudin-5 were stained in red. The cell nucleus was stained in blue with DAPI. Scale bar, 10 μm.

Article Snippet: ZO-1 (ab216880, 1:1000) and Claudin-5 (ab131259, 1:2000) antibodies were obtained from Abcam (Cambridge, MA). β-actin (66009-1-Ig, 1:50000), Occludin (13409-1-AP, 1:2000), VEGFA (19003-1-AP, 1:1000), and STAT3 (60199-1-Ig, 1:5000) antibodies were obtained from Proteintech (Chicago, IL). p-STAT3 (9145S, 1:2000) was obtained from Cell Signaling Technology (Danvers, MA).

Techniques: CRISPR, Knock-Out, Amplification, Western Blot, Expressing, Control, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: Cytokine MIF Enhances Blood-Brain Barrier Permeability: Impact for Therapy in Ischemic Stroke

doi: 10.1038/s41598-017-16927-9

Figure Lengend Snippet: Administration of exogenous MIF increases and ISO-1 inhibits the BBB permeability following tMCAO in rats. Evans blue dye was used to evaluate the BBB permeability in vivo . ( A ) The time-course of BBB permeability assay. ( B ) Representative images of Evans blue dye in brain tissue. Note that Evans blue dye increased in ipsilateral cortex following tMCAo. MIF increased and ISO-1 inhibited stroke-induced increase of BBB permeability. ( C ) The summarized results of dye content in the cortex tissue. ( D ) The expressions of GFAP and tight junction proteins occludin, claudin-5 and ZO-1 in brain tissue (full-length blots are presented in Supplementary Figure ). Data are presented as mean ± S.E.M. (n = 4–6 in Evans blue assay, and n = 5 in Western blotting). * p < 0.05 compared with contralateral cortex. # p < 0.05 compared with control group.

Article Snippet: Primary antibodies: occludin (1:1000; Santa Cruz Biotechnology, CA, USA), claudin-5 (1:500, Aviva systems Biology, CA, USA), ZO-1 (1:1000; Invitrogen, CA, USA), GAPDH (1:10000; Santa Cruz Biotechnology, CA, USA), GFAP (1:1000; Santa Cruz Biotechnology, CA, USA).

Techniques: Permeability, In Vivo, Evans Blue Assay, Western Blot, Control

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 2. IL-22 promotes BBB permeability enhancement in vivo. (A) Effects of the recombinant mouse IL-22 (10 ng/mouse) on the permeability of the mice brain evaluated using the Evans blue approach (n = 5). (B) Western blot analysis of ZO-1, Occludin, and Claudin-5 in the brains of IL- 22-treated (10 ng/mouse) mice (n = 3). β-actin was used as the loading control. Densitometry was performed to analyze differences among the samples. **p < 0.01, ***p < 0.001. (C) Brain samples of both mice treated with and without IL-22 (10 ng/mouse) for 24 h were analyzed for vascular endothelium integrity by immunofluorescence. The figures represent the results of three mouse brains (three images per mouse). ZO-1, Occludin, and Claudin-5 were selected as the markers reflecting the vascular endothelium integrity. CD31 was specifically applied for labeling the microvessels in green. The cell nucleus was stained in blue with DAPI. Scale bar indicates 50 μm.

Article Snippet: The sections were then incubated with a primary CD31 antibody (Proteintech, 66065-2-Ig, 1:100), followed by incubation with the appropriate secondary antibody, fluorescein isothiocyanate (Proteintech, SA00003-9, 1:100), prior to final nuclear staining with DAPI.

Techniques: Permeability, In Vivo, Recombinant, Western Blot, Control, Immunofluorescence, Labeling, Staining

Journal: ACS infectious diseases

Article Title: Interleukin-22 Contributes to Blood-Brain Barrier Disruption via STAT3/VEGFA Activation in Escherichia coli Meningitis.

doi: 10.1021/acsinfecdis.3c00668

Figure Lengend Snippet: Figure 3. IL-22 knockout in mice suppresses meningitic E. coli-induced BBB disruption. (A) Evans blue assay was used to evaluate the permeability of WT and IL-22−/−mice with or without meningitic E. coli infection (n = 5). (B) Brain lysates from the challenged WT and IL-22−/−mice were assessed using Western blot for the expression of TJPs (n = 3). β-Actin was used as the loading control. Densitometry was performed to analyze differences among the samples. *p < 0.05, **p < 0.01, ***p < 0.001. (C) Immunofluorescence analysis of vascular endothelium integrity in infected WT and IL-22−/−mice. The figures represent the results of three mouse brains (three images per mouse). ZO-1, Occludin, and Claudin-5 were stained in red. CD31 was specifically applied for labeling the microvessels in green. The cell nucleus was stained in blue with DAPI. Scale bar indicates 50 μm.

Article Snippet: The sections were then incubated with a primary CD31 antibody (Proteintech, 66065-2-Ig, 1:100), followed by incubation with the appropriate secondary antibody, fluorescein isothiocyanate (Proteintech, SA00003-9, 1:100), prior to final nuclear staining with DAPI.

Techniques: Knock-Out, Disruption, Evans Blue Assay, Permeability, Infection, Western Blot, Expressing, Control, Immunofluorescence, Staining, Labeling

Journal: Scientific Reports

Article Title: Cytokine MIF Enhances Blood-Brain Barrier Permeability: Impact for Therapy in Ischemic Stroke

doi: 10.1038/s41598-017-16927-9

Figure Lengend Snippet: Administration of exogenous MIF increases and ISO-1 inhibits the BBB permeability following tMCAO in rats. Evans blue dye was used to evaluate the BBB permeability in vivo . ( A ) The time-course of BBB permeability assay. ( B ) Representative images of Evans blue dye in brain tissue. Note that Evans blue dye increased in ipsilateral cortex following tMCAo. MIF increased and ISO-1 inhibited stroke-induced increase of BBB permeability. ( C ) The summarized results of dye content in the cortex tissue. ( D ) The expressions of GFAP and tight junction proteins occludin, claudin-5 and ZO-1 in brain tissue (full-length blots are presented in Supplementary Figure ). Data are presented as mean ± S.E.M. (n = 4–6 in Evans blue assay, and n = 5 in Western blotting). * p < 0.05 compared with contralateral cortex. # p < 0.05 compared with control group.

Article Snippet: Primary antibodies: occludin (1:1000; Santa Cruz Biotechnology, CA, USA), claudin-5 (1:500, Aviva systems Biology, CA, USA), ZO-1 (1:1000; Invitrogen, CA, USA), GAPDH (1:10000; Santa Cruz Biotechnology, CA, USA), GFAP (1:1000; Santa Cruz Biotechnology, CA, USA).

Techniques: Permeability, In Vivo, Evans Blue Assay, Western Blot