anti-cd44 [im7] anti-mouse, apc-efluor 780  (Thermo Fisher)

Journal: Nature Communications

Article Title: Distinctive CD39 + CD9 + lung interstitial macrophages suppress IL-23/Th17-mediated neutrophilic asthma by inhibiting NETosis

doi: 10.1038/s41467-024-53038-2

Figure Lengend Snippet: a Schematic diagram of allergen sensitization and challenge protocol. In the NDA mouse model, sensitization with PBS or OVA + LPS intranasally ( i.n .) was followed by challenge with PBS or OVA ( i.n .) at the indicated days. Isotype control, αIL-23p19, or dexamethasone (Dex), was treated at 1 hr before every challenge period. The inflammatory response was assessed at 48 hr after the last challenge. b Total cell, neutrophil, and eosinophil counts in BALF measured using flow ( n = 8 mice per group). c RORγt + , T-bet + , and GATA3 + Th cell counts in CD4 + T cells (Live,Dump - FOXP3 - CD44 + CD4 + TCRb + ) measured using flow cytometry ( n = 6 mice per group). d Levels of Th17 (IL-17), Th1 (IFN-γ), and Th2 (IL-13) cell-related cytokine in BALF ( n = 8 mice per group). e Representative H&E staining of lung sections obtained from the indicated conditions. Scale bar = 100 μm. f Inflammatory scores quantified from H&E staining (PBS/PBS ( n = 3), OVA + LPS/PBS ( n = 3), OVA + LPS/OVA ( n = 3), OVA + LPS/OVA + αIL-23p19 ( n = 5), and OVA + LPS/OVA+Dex ( n = 5)). g Representative PAS staining of lung sections obtained from the indicated conditions. Scale bar = 200 μm. h PAS-positive area quantified from PAS staining ( n = 4 mice per group) ( i )P enh values measured using whole body plethysmography (PBS/PBS ( n = 8), OVA + LPS/PBS ( n = 9), OVA + LPS/OVA ( n = 8), OVA + LPS/OVA + αIL-23p19 ( n = 5), and OVA + LPS/OVA+Dex ( n = 8)). j R rs values measured using an invasive ventilated lung resistance method (PBS/PBS ( n = 3), OVA + LPS/PBS ( n = 4), OVA + LPS/OVA ( n = 5), OVA + LPS/OVA + αIL-23p19 ( n = 4), and OVA + LPS/OVA + Dex ( n = 5)). Data are shown as mean ± s.e.m. Significance was determined by one-way analysis of variance (ANOVA) with Tukey’s post hoc correction ( b – d , f , h ) and two-way ANOVA with Sidak’s post hoc correction ( i , j ).

Article Snippet: To examine the T cell population, the lung single-cell samples were stained with LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (L34957, ThermoFisher), Percp/Cyanine 5.5-anti-mouse/human CD11b (M1/70, Biolegend), Percp/Cyanine 5.5-anti-CD14 (45-0141-82, Sa2-8, eBioscience), Percp/Cyanine 5.5-anti-CD19 (115533, 6D5, Biolegend), Percp/Cyanine 5.5-anti-B220 (45-0452-82, RA3-6B2, eBioscience), Percp/Cyanine 5.5-anti-I-A/1-E (107625, M5/114.15.2, Biolegend), Brilliant Violet 605-anti-mouse TCRβ chain (109241, H57-597, Biolegend), BV421 anti-mouse RORγt (562894, Q31-378, BD), Alexa Fluor 488-anti-FOXP3 (53-5773-82, FJK-16s, eBioscience), PE-CF594-anti-T-bet (562467, O4-46, BD), APC-eFluor-780 CD44 (47-0441-82, IM7, eBioscience), Brilliant Violet-711 anti-mouse CD4 (100557, RM4-5, Biolgend), and PE-anti-GATA3 (12-9966-42, TWAJ, eBioscience).

Techniques: Control, Flow Cytometry, Staining

Journal: Molecular Pain

Article Title: Regulatory T-cells and IL-5 mediate pain outcomes in a preclinical model of chronic muscle pain

doi: 10.1177/17448069221110691

Figure Lengend Snippet: Antibodies used in flow cytometry experiments.

Article Snippet: Anti-CD44 APC eFluor 780 conjugate , eBioscience , 47-0441-82 , 1:100 , 1 only.

Techniques: Flow Cytometry, Purification