Journal: Scientific reports

Article Title: Curcumin enhances elvitegravir concentration and alleviates oxidative stress and inflammatory response.

doi: 10.1038/s41598-023-47226-1

Figure Lengend Snippet: Figure 3. Western blot analysis of neural protein markers upon treatment with EVG (25 mg/kg), and CUR (20 mg/kg) in Balb/c mice brain. Protein was isolated from treated mice and expression of (a) NeuN, (b) Synaptophysin, (c) L1CAM, (d) GFAP was analyzed using western blotting. Results are expressed as means ± S.E.M (n = 4). Statistical analysis was performed using one-way ANOVA with Tukey’s post-hoc test for multiple group comparisons.

Article Snippet: The primary antibodies used were SOD1 mouse monoclonal antibody (1:1500 dilution, catalog no. sc-101523, Santa Cruz Biotechnology), CAT mouse monoclonal antibody (1:1000 dilution, catalog no. sc-365738, Santa Cruz Biotechnology), IL-1β rabbit polyclonal antibody (1:1000 dilution, catalog no. 16806-1AP, Proteintech), TNF-α rabbit polyclonal antibody (1:1000 dilution, catalog no. 16806-1AP, Proteintech), β-Actin rabbit polyclonal antibody (1:4000 dilution, catalog no. 20536-1-AP, Proteintech), NeuN rabbit polyclonal antibody (1:1000 dilution, catalog no. 26975-1-AP, Proteintech), Synaptophysin mouse monoclonal antibody (1:20,000 dilution, catalog no. 67864-1-Ig, Proteintech), β-Actin mouse monoclonal antibody (1:20,000 dilution, catalog no. 66009-1-Ig, Proteintech), GFAP rabbit polyclonal antibody (1:1000 dilution), and L1CAM rabbit polyclonal antibody (catalog no. 20659-1-AP, Proteintech Inc, Rosemont, IL, USA).

Techniques: Western Blot, Isolation, Expressing

Journal: bioRxiv

Article Title: Neuron-dependent tuft cell expansion initiates sinonasal allergic Type 2 inflammation

doi: 10.1101/2023.07.04.547596

Figure Lengend Snippet: (A-G) Data from Pou2f3-/- mice and litter mate controls treated with PBS or FAM, percentages of (A) Tuft cells 1 and 2, (B) total and Ki67+ basal cells, (C) ILC2, (D) CD4+ TH2 cells, (E) eosinophils, numbers and percentages of (F) M2 macrophages, and of (G) cDC2. (H) Protocol for treatment of Advillin-Cre;R26-DTR– and Advillin-Cre;R26-DTR+ mice given 100ng of diptheria toxin (DT) i.n. one day prior to the start of i.n. FAM challenge and on day D7 with analysis on D12. (I) Immunostaining of DCLK1+ STC (green, yellow asterisks) and PGP9.5+ neuron afferents (red) in the respiratory epithelium of Advillin-Cre;R26-DTR– (lacking DTR expression, left) and Advillin-Cre;R26-DTR+ (expressing DTR, right) mice following both DT treatments combined with FAM challenge. scale bars = 100μm. (J) Immunostaining for synaptophysin+ afferents (red) in the respiratory epithelium of DT-treated Advillin-Cre+;R26-DTR- (left) and Advillin-Cre+;R26-DTR- (right) mice that did not receive i.n. FAM, two representative images per genotype, scale bars = 50 μm. (K-L) Numbers and percentages of (K) Tuft-1 and (L) Tuft-2 cells. (M) Levels of STC products IL-25 (left) and CysLT (right). (N-Q) Numbers and percentages of (N) ILC2, (O) M2 Macrophages, (P) eosinophils, and (Q) CD4+ TH2 cells. (R) Sneeze-like events evoked by i.n. capsaicin (12µM), determined from 10-minute video recordings with sound. (S) Levels of neuropeptides SP and NMU in the sinonasal wash. Data are mean ± SEM, representative of 3-4 experiments (4-5 mice/treatment), one-way ANOVA with Tukey’s post-hoc tests. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Article Snippet: The primary antibodies used for immunofluorescence staining are rabbit anti-mouse DCAMLK1 antibody (1:1000, Abcam), chicken anti-mouse Keratin 5 Polyclonal Antibody (1:1000; BioLegend), anti-mouse Nav1.7 Antibody (1:500, Novus Biologicals), rabbit anti-mouse Synaptophysin Polyclonal antibody (1:1000, Proteintech), anti-mouse PGP9.5 Polyclonal Antibody (1:1000, ThermoFisher), guinea pig anti-mouse TRPV1 Polyclonal Antibody (1:500, ThermoFisher), and DAPI Solution (1:1000; ThermoFisher).

Techniques: Immunostaining, Expressing

Journal: bioRxiv

Article Title: Neuron-dependent tuft cell expansion initiates sinonasal allergic Type 2 inflammation

doi: 10.1101/2023.07.04.547596

Figure Lengend Snippet: (A) Immunofluorescence staining of naïve respiratory epithelium for DCLK1+, Keratin-5+, and Synaptophysin+ lining the airway (AW). Yellow arrows point to neuron afferents in contact with the airway and pink arrow points to neuron afferents in proximity to STC, scale bar = 100μm. (B-C) Sinonasal wash fluid levels of (B) substance P and (C) neuromedin U in mice exposed to PBS or at D5 vs. D12 post FAM exposure. (D) Capsaicin (12µM i.n.) evoked sneezing-like behavior, which was counted from 10-minute sound and video recordings in a blinded manner at D5 vs. D12 post FAM exposure. (E) Protocol for exposing mice to 100ng of RTX to deplete TRPV1 afferents in trigeminal ganglia. (F) Sinonasal fluid levels of SP (left) or NMU (right) neuropeptides at the indicated timepoints. (G) Sinonasal levels of IL-25 (left) and CysLT (right). (H-O) Total number of (H) Tuft-1 cells, (I) Tuft-2 cells, (J) basal cells, (K) Eosinophils, (L) cDC2s, (M) ILC2s, (N) IL-5+(left) and IL-13+ (right) ILC2s, and (O) CD4+ TH2 cells in respiratory epithelium. Data are mean ± SEM, representative of 3-4 experiments (4-5 mice/treatment), one-way ANOVA with Tukey’s post-hoc tests. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001. See also and .

Article Snippet: The primary antibodies used for immunofluorescence staining are rabbit anti-mouse DCAMLK1 antibody (1:1000, Abcam), chicken anti-mouse Keratin 5 Polyclonal Antibody (1:1000; BioLegend), anti-mouse Nav1.7 Antibody (1:500, Novus Biologicals), rabbit anti-mouse Synaptophysin Polyclonal antibody (1:1000, Proteintech), anti-mouse PGP9.5 Polyclonal Antibody (1:1000, ThermoFisher), guinea pig anti-mouse TRPV1 Polyclonal Antibody (1:500, ThermoFisher), and DAPI Solution (1:1000; ThermoFisher).

Techniques: Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Neuron-dependent tuft cell expansion initiates sinonasal allergic Type 2 inflammation

doi: 10.1101/2023.07.04.547596

Figure Lengend Snippet: (A) Immunostaining of synaptophysin+ nerves (red) lining the airway (AW) of the respiratory epithelium following D5 FAM to assess the effect of DT-treatment in DTR-(left) or DTR+ (right) TRPV1-Cre+ mice on general innervation (top and bottom images from separate mice), scale bar = 50 µm. (B) Capsaicin (12µM i.n.) was used to evoke sneezing-like behavior, which was counted from sound and video recording in a treatment-blind manner. (C-G) The percentages of (C) Tuft-1, (D) Tuft-2, (E) total basal cells, (F) eosinophils, and (G) cDC2 in the respiratory epithelium as determined by flow cytometry. (H-K) The percentage of (H) total ILC2, number and percentages of (I) IL-5+ or IL-13+ ILC2, (J) M2 Macrophages, and (K) percentages of CD4+ TH2 cells in the respiratory epithelium at D5 or 12. Data are mean ± SEM, representative of 2-3 experiments (4-5 mice/treatment), one-way ANOVA with Tukey’s post-hoc tests. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Article Snippet: The primary antibodies used for immunofluorescence staining are rabbit anti-mouse DCAMLK1 antibody (1:1000, Abcam), chicken anti-mouse Keratin 5 Polyclonal Antibody (1:1000; BioLegend), anti-mouse Nav1.7 Antibody (1:500, Novus Biologicals), rabbit anti-mouse Synaptophysin Polyclonal antibody (1:1000, Proteintech), anti-mouse PGP9.5 Polyclonal Antibody (1:1000, ThermoFisher), guinea pig anti-mouse TRPV1 Polyclonal Antibody (1:500, ThermoFisher), and DAPI Solution (1:1000; ThermoFisher).

Techniques: Immunostaining, Flow Cytometry