neuron specific beta iii tubulin nl557 conjugated antibody  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: NF-κB/IL-6 axis drives impaired corneal wound healing in aqueous-deficient dry eye

doi: 10.3389/fimmu.2025.1684290

Figure Lengend Snippet: LGE impairs corneal nerve regeneration and sensory recovery following epithelial abrasion. (A) Representative immunofluorescence images of corneal nerve fibers at 0, 2, 4, and 6 days after epithelial abrasion in control and LGE‐treated mice. Corneal nerves were visualized using an anti‐β‐III tubulin antibody conjugated with NL557. Upper panels show wide‐field views (scale bars = 500 μm); lower panels show magnified views of the central cornea (scale bars = 200 μm). (B) Quantification of corneal nerve density in the central 1500 μm × 1500 μm region using Imaris 6.2 software (as illustrated in Supplementary Figure 3 ). (C) Representative images of retraction bulbs in the central cornea at 6 hours post‐abrasion (scale bars = 40 μm). (D) Quantification of retraction bulb numbers in control and LGE‐treated mice. (E) Corneal mechanical sensitivity measured using a Cochet‐Bonnet esthesiometer at days 1–6 post‐abrasion. LGE‐treated mice exhibited reduced corneal sensitivity compared to controls at days 3–6. All data are presented as mean ± SD. Sample size: n = 6 mice per group (B, D, E) . Statistical significance: * P < 0.05, and ** P < 0.01.

Article Snippet: Subsequently, whole corneas were systematically incubated overnight at 4°C with one of the following standardized primary antibodies: i) Anti‐mouse Ly6G conjugated with fluorescein isothiocyanate (FITC; 1:100 dilution; BD Biosciences, Cat. No. 551460); ii) Anti‐mouse GL3 conjugated with phycoerythrin (PE; 1:100 dilution; BD Biosciences, Cat. No. 553178); iii) Anti‐β‐III tubulin conjugated with NL557 (1:10 dilution; R&D Systems, Cat. No. NL1195R).

Techniques: Immunofluorescence, Control, Software

Journal: Frontiers in Immunology

Article Title: NF-κB/IL-6 axis drives impaired corneal wound healing in aqueous-deficient dry eye

doi: 10.3389/fimmu.2025.1684290

Figure Lengend Snippet: Effects of NF-κB or IL-6 inhibition on corneal re‐epithelialization and nerve regeneration in LGE-treated mice. (A) Representative fluorescein sodium staining images of corneal epithelial wounds at multiple time points (0–42 hours) after abrasion in control mice, and LGE‐treated mice administered PBS, CAPE, isotype IgG, or anti‐IL‐6 antibody. (B, C) Quantification of residual corneal wound area over time in each group of mice. (B) Comparison among control, LGE + PBS, and LGE + CAPE groups. (C) Comparison among control, LGE + isotype IgG, and LGE + anti‐IL‐6 antibody groups. (D, E) Quantification of dividing basal epithelial cells in each group of mice. (D) Comparison among control, LGE + PBS, and LGE + CAPE groups; (E) Comparison among control, LGE + isotype IgG, and LGE + anti‐IL‐6 antibody groups. (F) Immunofluorescence images of whole-mount corneal nerves at 6 days post‐injury using anti‐β‐III tubulin antibody conjugated with NL557. Upper panels show full corneal view (scale bars = 500 μm); lower panels show magnified central region (scale bars = 200 μm). (G, H) Quantification of corneal nerve fiber density (G) and mechanical sensitivity (H) in control, LGE + PBS, and LGE + CAPE groups. (I, J) Quantification of corneal nerve fiber density (I) and mechanical sensitivity (J) in control, LGE + isotype IgG, and LGE + anti‐IL‐6 antibody groups. All values are presented as mean ± SD. Sample size: n = 6 mice in each group (B–E, G–J) . Statistical significance: * P < 0.05, ** P < 0.01, and *** P < 0.001 for LGE‐treated mice versus control mice; # P < 0.05, ## P < 0.01, and ### P < 0.001 for LGE + CAPE or anti‐IL‐6 antibody versus respective vehicle (PBS or IgG); + P < 0.05, ++ P < 0.01, and +++ P < 0.001 for LGE + CAPE or anti‐IL‐6 antibody versus control.

Article Snippet: Subsequently, whole corneas were systematically incubated overnight at 4°C with one of the following standardized primary antibodies: i) Anti‐mouse Ly6G conjugated with fluorescein isothiocyanate (FITC; 1:100 dilution; BD Biosciences, Cat. No. 551460); ii) Anti‐mouse GL3 conjugated with phycoerythrin (PE; 1:100 dilution; BD Biosciences, Cat. No. 553178); iii) Anti‐β‐III tubulin conjugated with NL557 (1:10 dilution; R&D Systems, Cat. No. NL1195R).

Techniques: Inhibition, Staining, Control, Comparison, Immunofluorescence

Journal: ACS nano

Article Title: Combination Nanomedicine Strategy for Preventing High-Risk Corneal Transplantation Rejection.

doi: 10.1021/acsnano.4c06595

Figure Lengend Snippet: Figure 7. Corneal nerve regeneration after HR PKP surgery. (A) Immunostaining images of corneal nerve marker β-tubulin III, scale bar 200 μm. (B) Corneal nerve regeneration was quantified as the percentage of threshold area positive for β-tubulin III staining in the center part of cornea with the diameter of 3.5 mm. Statistical analysis for B: one-way ANOVA with a Tukey post hoc test for multiple comparison. (*p ≤0.05; **p ≤0.01; and ***p ≤0.001). N = 3 for each group. All data are plotted from mean ± SEM.

Article Snippet: After that, cornea samples were washed with PBS with 0.3% Triton X-100 (Sigma-Aldrich) for 4 × 10 min followed by blocking with 2% BSA-PBS-Triton X-100 solution at room temperature for another 2 h. The cornea was incubated at 4 °C overnight in fluorescein-labeled antirat β-tubulin III antibody (R&D systems, Cat No: NL1195R).

Techniques: Immunostaining, Marker, Staining, Comparison