Fig 1: Inhibiting HK2 activity by 2-DG protected mice from airway inflammation. (A,B) Images and statistical graph (n = 6) of lung histology. (C) BALF total cells counts. (D) Differential counts of inflammatory cells in the BALF (n = 6). (E) mRNA levels of IL-4, IL-13, IL-33 and CCL20 in lungs of OVA-immunized mice treated with 2-DG or not (n = 6 per group). (F) AHR, including Rrs, Errs, and Crs, was recorded 24 h after the last flexiVent challenge. (n = 6). Elevated HK2 expression in airway epithelial cells regulates airway epithelial cells glycolysis, airway inflammation and cell death, contributing to the pathological of asthma. AHR = airway hyperresponsiveness, Rrs = respiratory system resistance; Ers = respiratory system elastance; Crs = respiratory system compliance. The data were shown as mean ± SEM, statistical differences were determined with one-way ANOVA or two-way ANOVA with a Tukey-Kramer test for comparison. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Fig 2: HK2 is upregulated in asthma. (A) Representative images showing HK2 protein immunostaining (brown color) in patients with asthma (n = 7) compared with that seen in healthy subjects (n = 8), HK2 protein relative expression was analyzed by Image-pro plus 6.0 and presented graphically. The images were captured under an original magnification of ×400. Scale bar, 100 μm. (B) Western blot analysis of HK2 in the lungs of experimental animals (n = 6 per group). (C) RT-PCR analysis of HK2 in the lungs of experimental animals (n = 6 per group). (D) Representative results of co-immunostaining of Scgb1a1 (secretoglobin family 1A member 1) and HK2 in the lung sections of experimental animals. The images were captured under an original magnification of ×200. Scale bar, 50 μm. (E) HK2 mRNA levels were measured after 48 h of stimulation with HDM, IL-1β, IL-33, IL-6, IL-8, LPS, TGF-β, or TNF-α (n = 6 per group). (F) Western blot results demonstrating HK2 expression in Beas-2B cells following treatment with IL-1β (n = 4 per group). Data are presented as mean ± SEM, Statistical differences were determined with a student’s t test for comparison between the two groups and one-way ANOVA with a Tukey-Kramer test for multiple-group comparisons. * p < 0.05, ** p < 0.01, **** p < 0.0001.
Fig 3: IL-33/ST2 axis promotes angiogenesis in melanoma in vivo.a Schematic view of the treatment that C57BL/6 mice burdened with B16F10 melanoma cell tumors with or without mouse IL-33 protein or A-485 treatment. b Images of B16F10 melanoma cell xenografts isolated from mice with indicated treatments. Tumor volumes, tumor weights, and mouse weights in each group were calculated and displayed in (c, d, e) (n = 5). f 4D-OCT intravital imaging of microvasculature in tumor with indicated treatments (n = 5, Scale bar, 300 μm). g, h Immunofluorescence staining of CD31, ST2 and H3K18la in tumor with or without IL-33 mouse protein or A-485 treatment (n = 3, Scale bar, 100 μm). P value was calculated by one-way ANOVA followed by Tukey’s multiple comparisons test, mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001. ns non-significant.
Fig 4: IL-33/ST2 axis promotes angiogenesis in melanoma in vitro.a Relative mRNA level of ST2 in TA-HUVEC with or without the knockdown of ST2. b Immunoblotting analysis of ST2 expression in TA-HUVEC with or without the knockdown of ST2. c Cell viability of TA-HUVEC with or without ST2 knockdown, treated with IL-33 (10 ng/ml), NALA (10 mM) and/or A-485 (10 μM) for 48 h. d Cell migration of TA-HUVEC with or without ST2 knockdown, treated with IL-33 (10 ng/ml), NALA (10 mM) and/or A-485 (10 μM) for 48 h. Scale bar, 200 μm. e Tube formation of TA-HUVEC with or without ST2 knockdown, treated with IL-33 (10 ng/ml), NALA (10 mM) and/or A-485 (10 μM) for 48 h. Scale bar, 200 μm. P-value was calculated by one-way ANOVA followed by Tukey’s multiple comparisons test, mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001. ns non-significant.
Supplier Page from MedChemExpress for IL-33 Protein, Human