Journal: The Journal of allergy and clinical immunology

Article Title: A Jagged 1–Notch 4 molecular switch mediates airway inflammation induced by ultrafine particles

doi: 10.1016/j.jaci.2018.03.009

Figure Lengend Snippet: AM supports UFP-dependent upregulation of Th cell cytokine production by allergen-specific CD4+ T cells in a Notch4-dependent manner. A, Representative flow cytometric analysis of IL-4, IL-13 and IL-17 cytokine production by naive Il4raR576CD4+DO11.10+ T cells co-cultured with FACS-purified AM isolated from l4raR576 or Il4raR576Lyz2CreJag1Δ/Δ mice that have been pulsed with OVA323-339 peptide in the presence of UFP (10 μg/mL). Co-cultures were treated with either isotype control (Iso) Ab or an anti-Notch4 mAb, as indicated, and cytokine analysis was carried out on gated CD4+Foxp3− T cell. B, Frequencies of T cells expressing the respective cytokine upon co-culture with AM that have been either sham treated (PBS) or pulsed with OVA323-339 peptide alone or in combination with UFP (10 μg/mL). Anti-Notch4 mAb or isotype control Ab were added as indicated. Results are representative of 3 independent experiments. *P < .05, **P < .01, ***P < .001, and ****P < .0001, two-way ANOVA with post-test analysis.

Article Snippet: For anti-Notch4 antibody blocking, 150 μg Armenian hamster anti-mouse Notch4 IgG mAb (clone HMN4-14; Bio X Cell) 35 , or control Armenian hamster IgG polyclonal antibodies (Ab) (Bio X cell), were suspended in 100 μl PBS buffer and administered daily for three consecutive days during OVA aerosol challenge.

Techniques: Cell Culture, Purification, Isolation, Expressing, Co-Culture Assay

Journal: The Journal of allergy and clinical immunology

Article Title: A Jagged 1–Notch 4 molecular switch mediates airway inflammation induced by ultrafine particles

doi: 10.1016/j.jaci.2018.03.009

Figure Lengend Snippet: UFP enhances allergic airway inflammation in a Notch4-dependent manner. A, Representative PAS staining of lung tissues isolated from Il4raR576 mice sensitized and challenged with OVA alone, or together with UFP, in the presence of either isotype control (Iso) Ab or an anti-Notch4 mAb. B, Inflammation scores in lung tissues of the mouse groups described in in Fig 7, A. C–H Airway hyper-responsiveness in response to methacholine (Fig 7, C), absolute numbers of eosinophils (Fig 7, D), T cells (Fig 7, E) and neutrophils (Fig 7, F) in the BAL fluids, total (Fig 7, G) and OVA-specific (Fig 7, H) levels in the serum of the mouse groups described in Fig 7, A. I-L, Absolute numbers of lung Foxp3−CD4+T cells secreting IL-4 (Fig 7, I), IL13 (Fig 7, J), IL-17 (Fig 7, K) and IFN-γ (Fig 7, L) in the mouse groups described in Fig 7, A. M–P, Absolute numbers of lung Foxp3+CD4+Treg cells secreting IL-4 (Fig 7, M), IL13 (Fig 7, N), IL-17 (Fig 7, O) and IFN-γ (Fig 7, P) in the mouse groups described in panel Fig 7, A. Results are representative of 2 independent experiments. N=5 mice/group. *p<0.05, **<0.01, ***<0.001, ****<0.0001 by two-way ANOVA with post test analysis.

Article Snippet: For anti-Notch4 antibody blocking, 150 μg Armenian hamster anti-mouse Notch4 IgG mAb (clone HMN4-14; Bio X Cell) 35 , or control Armenian hamster IgG polyclonal antibodies (Ab) (Bio X cell), were suspended in 100 μl PBS buffer and administered daily for three consecutive days during OVA aerosol challenge.

Techniques: Staining, Isolation

Journal: International Journal of Oncology

Article Title: Regulation of gliomagenesis and stemness through acid sensor ASIC1a

doi: 10.3892/ijo.2021.5262

Figure Lengend Snippet: Notch signaling is negatively associated with ASIC1a in glioblastoma multiforme. (A-C) The glioma cells were transfected with shASIC1a and control followed by assaying protein expression of ASIC1a, active form of Notch1, Notch2, Notch3, Notch4, and Notch target survivin, as well as GSC markers CD133 and ALDH1 by western blotting in (A) A172, (B) U87MG, and (C) PDX-L12 cells. (D-F) Subsequently, the glioma cells (D) A172, (E) U87MG and (F) PDX-L12 were transfected with ASIC1a-FLAG followed by assaying protein expression of ASIC1a, active form of Notch1, Notch2, Notch3, Notch4 and Notch target survivin, as well as GSC markers CD133 and ALDH1 by western blotting. * P<0.05 and ** P<0.01. ASIC1a, acid-sensing ion channel 1a; sh, short hairpin; GSC, glioblastoma stem cells; ALDH1, aldehyde dehydrogenase 1.

Article Snippet: The following primary antibodies were used in the present study: Rabbit monoclonal anti-ASIC1a antibody (cat. no. 35-156465) was purchased from American Research Products, Inc. Rabbit polyclonal anti-Notch4 antibody (cat. no. 07-189) and anti-β-actin antibody (product no. A3854) were purchased from Sigma-Aldrich; Merck KGaA.

Techniques: Transfection, Control, Expressing, Western Blot