PE anti-mouse CD122 (IL-2Rβ) 123210 from BioLegend

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Nature communications
July 28 2020

Fig 1: DETC development requires Skint2 and Skint1 which form heteromers.a qPCR analysis for Skint1 and Skint2 message in adult mouse ear epidermis normalised to Ppia. Control, n = 8, Skint2Δ/Δ, n = 8. Data are mean ± SD of a representative experiment of three independent experiments. b Analysis of E16.5 thymocytes in WT, Skint2Δ/Δ and Skint1Δ/Δ animals, gated on live γδ T cells (top panel). Thymic γδ cells (gate top row) were assessed for CD45RB, CD122, CD24 and CD62L and expression of the Vγ5 TCR. Left panel: WT vs. Skint2Δ/Δ, right panel: WT vs. Skint1Δ/Δ. c Quantification of cell populations in quadrants Q1 to Q4 as indicated in (b), normalised to mean of wt = 1 for each quadrant, mean ± SD. WT n = 20; Skint2Δ/Δ n = 5; Skint1Δ/Δ n = 8 (two-tailed Man–Whitney analysis). d FACS analysis of ear epidermis in control (Skint2Δ/+) and Skint2Δ/Δ mice. CD45+, TCRγδ+ cells (gate left panel) were stained for presence of Vγ5Vδ1+ DETC (stained by Vγ5 and 17D1 antibody, right panel). e Microscopy images of adult mouse ear epidermal sheets in control and Skint2Δ/Δ mice. Comparison of DETC stained for CD45 (top: blue) and Vγ5+ (middle: green) γδ-TCR(GL3+) cells (bottom: red). Scale bar 50 µm. f Quantification of DETC and Langerhans cell numbers from microscopy images, n = 15 for each genotype (data are from three independent experiments, two-tailed Man–Whitney analysis). g Immunoprecipitation of Flag-tagged Skint1 from FVB or NF-Skint1Tg animals. Left: scheme of FVB mice expressing Skint1 and Skint2. Scheme: Top: wt FVB mice express endogenous, untagged Skint1 and Skint2; bottom: NF-Skint1Tg animals express a Flag-tagged Skint1 and untagged Skint2 on the Skint1Tac background. Right: Immunoprecipitation with anti-Flag antibody from lysates of pooled thymi of FVB and NF-Skint1Tg animals (nFVB = 12, nNFSkint1Tg = 22, 1 experiment). Expression control in 293 lysates transduced with either Flag-Skint1 alone or Flag-Skint1 & HA-Skint2 constructs. Long arrows: Skint1 band, asterisks: non-specific bands reflecting anti-FLAG Ig chain detection. Full scans are provided in Supplementary Fig. 7.
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Cell
September 22 2016

Fig 2: A Gut IEL Selecting Element(A) Left: IEL composition in WT versus NU/NU mice; antibody GL2 detects TRDV2-2-encoded Vd4 chain. Right: surface CD122 expression on NU/NU V?7+ IELs (n = 12).(B) IEL composition (top), enumeration (bottom left), and CD122 expression (bottom right) in germ-free (GF), food antigen-free (FAF), or GF-FAF C57Bl/6 mice at weeks 9–13 (n = 4).(C) qRT-PCR of denoted genes.(D) Histological analysis of Btnl1 RNA (middle: RNAScope) and 3-hr BrdU incorporation in vivo (bottom) in paraffin-embedded SI gut rolls (n = 3).(E) RNAScope of Btnl1, Btnl4, and Btnl6 in WT versus Btnl1-/- and Btnl4-/- mice.Data are representative of two (B), two or more (D and E), or three or more (A) independent experiments. In (C), data are pooled from two independent experiments.All error bars represent mean ± SD. See also Figure S2.
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Cell
September 22 2016

Fig 3: Btnl1 Drives Selective Expansion and Maturation of Gut IEL(A) 3 hr EdU incorporation in vivo in ?d IEL subsets and thymocytes from week 4 WT versus Btnl1-/- mice (n = 9). (B) Surface phenotypes of V?7+ and V?7GL2+ IELs from week 3–5 WT and Btnl1-/- mice (n = 8).(C) Top: Thy1 and CD122 expression by V?7+ IEL from day 35 Btnl1-/- mice (n = 8). Bottom: surface phenotypes of CD122HIThy1- and CD122LOThy1+ V?7+ IELs from week 4–6 Btnl1-/- mice (n = 4). W, week.(D and E) IEL reconstitution and CD122 profiles in (D) irradiated TCRd-/- mice 9–10 weeks post-BM transfers from indicated donors (n = 7) and (E) irradiated CD45.2+ WT or Btnl1-/- mice 4–5 weeks post-BM transfers from CD45.1+ C57Bl/6 mice (n = 7).Data are representative of two (D and E) or three or more (C, top) independent experiments. Panels (A) and (C) (bottom) present data pooled from three or more experiments.All error bars represent mean ± SD. See also Figure S4.
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Nature communications
July 28 2020

Fig 4: Depletion of individual Btnl genes does not impact Vγ7 IEL numbers but differentially affects CD122 expression.a Top: Experimental scheme to analyse the effect of Btnl1 and Btnl6 tamoxifen-mediated depletion at different timepoints. Bottom: RNAscope analysis for Btnl1 and Btnl6 in animals of indicated genotypes at 3 or 17 days post tamoxifen administration. Data are representative micrographs from one time course experiment with numbers of gut sections stained per genotype as: day 8: nctrl: 4, nBtnl1f/f-VcreERT2+: 2, nBtnl6f/f-VcreERT2+: 4, day 22: nctrl: 4, nBtnl1f/f-VcreERT2+: 1, nBtnl6f/f-VcreERT2+: 2, scale bar: 200 µm. b Quantification of Vγ7+ cells at indicated timepoints post tamoxifen (red and blue side arrows denote for comparison the average percentage of Vγ7+ IEL in full knockout animals (see also Figs. 1–3). Data are mean ± SD, day 8: nctrl: 6, nBtnl1f/f-VcreERT2+: 2, nBtnl6f/f-VcreERT2+: 4, day 15: nctrl: 2, nBtnl1f/f-VcreERT2+: 3, nBtnl6f/f-VcreERT2+: 3, day 22: nctrl: 2, nBtnl1f/f-VcreERT2+: 1, nBtnl6f/f-VcreERT2+: 4. c Top: Experimental scheme to analyse the effect of Btnl1 and Btnl6 tamoxifen-mediated depletion after 3 days. Bottom: Vδ-chain usage in Vγ7+ IEL in control (black), Btnl1f/f,VillinCreERT2+ (orange), Btnl6Δgut (purple) and Btnl6f/f,VillinCreERT2+ (light blue) animals. The Vδ4 and Vδ6.3 chain gated on Vγ7+ IEL are plotted against each other. d Histogram of surface CD122 expression in indicated subpopulations of Vγ7+ IEL in animals of indicated genotypes. e Percentage of Vγ7 cells (left) and usage of the Vδ4 and Vδ6.3 chain (right) in Vγ7+ IEL, 56 days after tamoxifen in Btnl6f/f VillinCreERT2− (black) and Btnl6f/f VillinCreERT2+ (blue) animals. f Quantification of Vδ-chain usage in Vγ7+ IEL in control, Btnl1f/f,VillinCreERT2+ and Btnl6f/f,VillinCreERT2+ knockout animals, 3 (left graph) and 56 days (right) after tamoxifen administration. Mean ± SEM from two experiments per timepoint, left graph: nctrl: 6, nBtnl1f/f-VcreERT2+: 3, nBtnl6f/f-VcreERT2+: 6, nBtnl6Δgut: 2, right graph: nctrl: 5, nBtn61f/f-VcreERT2+: 15.
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Nature communications
July 28 2020

Fig 5: The intestinal IEL compartment is shaped by expression of distinct Btnl molecules.a Targeting strategy to generate conditional and constitutive Btnl1 and Btnl6 knockout mice. Depending on the Cre-transgenic strain used (i/ii) resulting animals are either ubiquitously deleted for the respective Btnl gene (Δ/Δ), or harbour a tissue-specific deletion (Δgut). Black: translated exons, grey: untranslated regions, blue triangles: loxP sites. b Quantification of Vγ7+ IEL (black gate in c) in Btnl1 and Btnl6-deficient strains. Data are mean ± SEM of ≥2 independent experiments. nctrl: 32, nBtnl1-KOMP: 4, nBtnl1Δgut: 8, nBtnl6Δgut: 8, nBtnl6ΔΔ: 9. Statistical analysis: Kruskal–Wallace and Dunn’s multiple comparison analysis. c FACS profiles of IEL preparations from animals of indicated genotypes, gated on TCRγδ+ cells. The label Btnl1Komp indicates germline Btnl1KO animals generated by the IMPC that have been described16. Black gate: all Vγ7+ cells, green gate: Vγ7+Vδ4+ cells. d Quantification of IEL: Vγ7+ IEL numbers (top), %Vγ7+ IEL (middle) and %CD122+ cells (bottom) in animals of indicated genotypes. Data are mean ± SD of a representative experiment. Top panel: nctrl: 12, nBtnl1: 2, nBtnl6Δgut: 2, nBtnl6Δ/Δ: 3. Statistical analysis: Kruskal–Wallace & Dunn’s multiple comparison. Middle and bottom panel: nctrl: 12, nBtnl1Δgut: 3, nBtnl6Δgut: 2, nBtnl6Δ/Δ: 3. Statistical analysis: Kruskal–Wallace & Dunn’s multiple comparison. e Histogram for surface expression of CD122 in Vγ7+ IEL from animals of the indicated genotypes. f Scheme depicting the strategy to generate Btnl146Indel mice. Short guide RNAs flanking the 5′ region of Btnl1 and 5′ region of Btnl6 were injected with HDR templates. Due to the nature of CRISPR/Cas9 the intervening region was excised and a Btnl146INDEL mouse lacking the Btnl1-4-6 locus was created. Blue triangle: loxP site that was inserted due to the nature of the HDR template (see Methods). g FACS analysis of TCRβ−CD3+ IEL in Btnl1/4/6-KO and Btnl146INDEL mice. Colours correspond to coloured bar graphs in (h) and (i). h Quantification of Vγ7+ IEL depicted in (g). Data are mean ± SEM of three independent experiments. nCtrl: 12, nBtnll1Δ/Δ: 7, nBtnl4KOMP: 7, nBtnl6Δgut: 11, nBtnl146Indel: 10. Statistical analysis: Kruskal–Wallace & Dunn’s multiple comparison. i Surface expression of CD122 (left) and Thy1.2 (right) in Vγ7+ IEL from animals of indicated genotypes.

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PE anti-mouse CD122 (IL-2Rβ)

BioLegend's PE anti-mouse CD122 (IL-2Rβ) is a Rat monoclonal antibody. The PE anti-mouse CD122 (IL-2Rβ) Antibody was generated using IL2RB as the antigen. It reacts with Mouse. This antibody has been shown to work in applications such as: Flow Cytometry.

Description

PE anti-mouse CD122 (IL-2Rβ) [TM-β1];

Product Specs
ItemPE anti-mouse CD122 (IL-2Rβ)
CompanyBioLegend
Price $214.00Supplier Page View Company Product Page
Catalog Number123210
NCBI Gene AliasesCD122, IL-15Rbeta, IL15Rbeta, Il-2/15Rbeta, Il-2Rbeta, p70
HostRat
Conjugate/TagPE
CloneTM-β1
ApplicationsFC - Quality tested
ReactivityMouse
Quantity100 µg
IsotypeRat IgG2b, kappa
Antibody TypePrimary
NCBI Full Gene Nameinterleukin 2 receptor, beta chain
Concentration0.2mg/ml
FormatPhosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
ImmunogenRat T cell line expressing mouse CD122 ( IL-2Rβ)
ClonalityMonoclonal
TargetCD122

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