nk1 1 pk136 pe cy7 Search Results


anti-nk1.1 pe-c7  (Thermo Fisher)
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Thermo Fisher anti-nk1.1 pe-c7
Anti Nk1.1 Pe C7, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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nk1.1-pe cy7  (Thermo Fisher)
90
Thermo Fisher nk1.1-pe cy7
Nk1.1 Pe Cy7, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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nk1 1 pk136 pe cy7  (Rockland Immunochemicals)
91
Rockland Immunochemicals nk1 1 pk136 pe cy7
Nk1 1 Pk136 Pe Cy7, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti mouse nk1 1 pk 136 pe cy7  (Thermo Fisher)
86
Thermo Fisher anti mouse nk1 1 pk 136 pe cy7
Study of ROS levels in NKT and CD4 T cells. (A) Freshly isolated thymocytes, splenocytes and liver lymphocytes were compared for the amount of ROS using 2’,7’-dichlorofluorescin diacetate (DCFDA) as described in Materials and Methods. NKT cells were identified by TCR-β low and CD1d-tetramer+. N= 8 mice per group. (B) Expression level of CD44, CD62L, and <t>NK1.1</t> was used to compare the amount of ROS in two subsets of splenic NKT cells. N=4 mice per group. (C) NKT, CD44+ and CD44− CD4 T cells, and NK cells from spleen and liver were compared for the levels of ROS. N=6 mice per group. (D) NKT and CD4 T cells from total splenocytes were compared for mitochondria-produced ROS using MitoSox, as described in the Methods. N=3 mice per group. (E) Gene expression of the members of the Nox gene family was assessed from sorted splenic NKT and CD4 T cells using qPCR. Gene expression was normalized to either β-actin or GAPDH. N=4 mice per group. All flow cytometric data were analyzed with live cells and C57BL/6 mice were used. One representative histogram is shown from at least three independent experiments. Error bars represent the mean ± SEM. **p <0.01; ***p<0.001; ****p<0.0001.
Anti Mouse Nk1 1 Pk 136 Pe Cy7, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pe-cy7 antimouse nk1.1  (Becton Dickinson)
90
Becton Dickinson pe-cy7 antimouse nk1.1
Study of ROS levels in NKT and CD4 T cells. (A) Freshly isolated thymocytes, splenocytes and liver lymphocytes were compared for the amount of ROS using 2’,7’-dichlorofluorescin diacetate (DCFDA) as described in Materials and Methods. NKT cells were identified by TCR-β low and CD1d-tetramer+. N= 8 mice per group. (B) Expression level of CD44, CD62L, and <t>NK1.1</t> was used to compare the amount of ROS in two subsets of splenic NKT cells. N=4 mice per group. (C) NKT, CD44+ and CD44− CD4 T cells, and NK cells from spleen and liver were compared for the levels of ROS. N=6 mice per group. (D) NKT and CD4 T cells from total splenocytes were compared for mitochondria-produced ROS using MitoSox, as described in the Methods. N=3 mice per group. (E) Gene expression of the members of the Nox gene family was assessed from sorted splenic NKT and CD4 T cells using qPCR. Gene expression was normalized to either β-actin or GAPDH. N=4 mice per group. All flow cytometric data were analyzed with live cells and C57BL/6 mice were used. One representative histogram is shown from at least three independent experiments. Error bars represent the mean ± SEM. **p <0.01; ***p<0.001; ****p<0.0001.
Pe Cy7 Antimouse Nk1.1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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nk1.1 (perpc- cyanine 5.5, bv395, or pe/cy7; pk136  (Thermo Fisher)
90
Thermo Fisher nk1.1 (perpc- cyanine 5.5, bv395, or pe/cy7; pk136
Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − <t>NK1.1</t> − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.
Nk1.1 (Perpc Cyanine 5.5, Bv395, Or Pe/Cy7; Pk136, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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nk1.1 pe-cy7 antibody  (Thermo Fisher)
90
Thermo Fisher nk1.1 pe-cy7 antibody
Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − <t>NK1.1</t> − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.
Nk1.1 Pe Cy7 Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti nk1 1 pe cy7  (Thermo Fisher)
86
Thermo Fisher anti nk1 1 pe cy7
Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − <t>NK1.1</t> − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.
Anti Nk1 1 Pe Cy7, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-mouse cd44 im7 percp-cy5.5  (Thermo Fisher)
90
Thermo Fisher anti-mouse cd44 im7 percp-cy5.5
Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − <t>NK1.1</t> − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.
Anti Mouse Cd44 Im7 Percp Cy5.5, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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nk1 1 pk136 pe cy7 ebioscience  (Thermo Fisher)
86
Thermo Fisher nk1 1 pk136 pe cy7 ebioscience
Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − <t>NK1.1</t> − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.
Nk1 1 Pk136 Pe Cy7 Ebioscience, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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nk 1.1 pe cy7 antibody  (Thermo Fisher)
90
Thermo Fisher nk 1.1 pe cy7 antibody
Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − <t>NK1.1</t> − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.
Nk 1.1 Pe Cy7 Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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741587 biotin anti mouse nk1 1  (Thermo Fisher)
99
Thermo Fisher 741587 biotin anti mouse nk1 1
Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − <t>NK1.1</t> − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.
741587 Biotin Anti Mouse Nk1 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Study of ROS levels in NKT and CD4 T cells. (A) Freshly isolated thymocytes, splenocytes and liver lymphocytes were compared for the amount of ROS using 2’,7’-dichlorofluorescin diacetate (DCFDA) as described in Materials and Methods. NKT cells were identified by TCR-β low and CD1d-tetramer+. N= 8 mice per group. (B) Expression level of CD44, CD62L, and NK1.1 was used to compare the amount of ROS in two subsets of splenic NKT cells. N=4 mice per group. (C) NKT, CD44+ and CD44− CD4 T cells, and NK cells from spleen and liver were compared for the levels of ROS. N=6 mice per group. (D) NKT and CD4 T cells from total splenocytes were compared for mitochondria-produced ROS using MitoSox, as described in the Methods. N=3 mice per group. (E) Gene expression of the members of the Nox gene family was assessed from sorted splenic NKT and CD4 T cells using qPCR. Gene expression was normalized to either β-actin or GAPDH. N=4 mice per group. All flow cytometric data were analyzed with live cells and C57BL/6 mice were used. One representative histogram is shown from at least three independent experiments. Error bars represent the mean ± SEM. **p <0.01; ***p<0.001; ****p<0.0001.

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Reactive Oxygen Species Regulate the Inflammatory Function of NKT Cells through Promyelocytic Leukemia Zinc Finger

doi: 10.4049/jimmunol.1700567

Figure Lengend Snippet: Study of ROS levels in NKT and CD4 T cells. (A) Freshly isolated thymocytes, splenocytes and liver lymphocytes were compared for the amount of ROS using 2’,7’-dichlorofluorescin diacetate (DCFDA) as described in Materials and Methods. NKT cells were identified by TCR-β low and CD1d-tetramer+. N= 8 mice per group. (B) Expression level of CD44, CD62L, and NK1.1 was used to compare the amount of ROS in two subsets of splenic NKT cells. N=4 mice per group. (C) NKT, CD44+ and CD44− CD4 T cells, and NK cells from spleen and liver were compared for the levels of ROS. N=6 mice per group. (D) NKT and CD4 T cells from total splenocytes were compared for mitochondria-produced ROS using MitoSox, as described in the Methods. N=3 mice per group. (E) Gene expression of the members of the Nox gene family was assessed from sorted splenic NKT and CD4 T cells using qPCR. Gene expression was normalized to either β-actin or GAPDH. N=4 mice per group. All flow cytometric data were analyzed with live cells and C57BL/6 mice were used. One representative histogram is shown from at least three independent experiments. Error bars represent the mean ± SEM. **p <0.01; ***p<0.001; ****p<0.0001.

Article Snippet: Flow cytometry assay The following antibodies were used: anti-mouse TCR-β (H57-597) APC or Pacific Blue, PBS57 loaded CD1d tetramer APC or Pacific Blue, anti-mouse CD4 (GK1.5) PerCp-Cy5.5, anti-mouse CD8 (53-6.7) Am-cyan, anti-mouse NK1.1 (PK-136) PE-Cy7, anti-mouse CD44 (IM7) PerCp-Cy5.5, anti-mouse CD69 (H1-2F3) PE-Cy7, anti-mouse CD62L (MEL-14) eVolve605, anti-mouse IFN-γ (XMG1.2) FITC or APC, anti-mouse IL-4 (11B11) PE-Cy7, anti-mouse IL-17 (TC11-18H10) APC-eFluor780, anti-T-bet (eBio4B10) FITC, anti-RORγt (AFKJS-9) Pacific Blue, anti-GATA3 (L50-823) APC, and anti-PLZF (Mags-21F7) PE (all from eBioscience).

Techniques: Isolation, Expressing, Produced

Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − NK1.1 − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Journal: The Journal of Experimental Medicine

Article Title: Amino acid availability acts as a metabolic rheostat to determine the magnitude of ILC2 responses

doi: 10.1084/jem.20221073

Figure Lengend Snippet: Metabolite and dietary factors influence innate type 2 response to helminth infection. (A) Relative levels of fecal amino acids and amino acid–related metabolites in control and day 7 post-infection (p.i.) N. brasiliensis –infected C57BL/6 mice ( n = 4 mice per group, representative of two independent experiments, data shows z -scores). (B) Relative abundance of selected amino acids in naive mice or mice infected with N. brasiliensis ( Nippo ; day 7 p.i., blue), H. polygyrus (day 7 p.i., purple), or T. spiralis (day 7 p.i., pink). n = 4 mice per group, representative of one experiment, data show relative abundance. (C–G) (C) Numbers of ILC2 in naive or N. brasiliensis –infected (day 7 p.i.) mice, (D) frequency and (E) number of Ki-67 + ILC2 on days 4 and 7 after N. brasiliensis infection, and (F) frequency and (G) number of IL-5 and IL-13 producing ILC2 on day 7 after N. brasiliensis infection in C57BL/6 mice fed a normal (21%) or low (5%) protein diet. n = 6–11 mice per group, pooled data from two independent experiments. ILC2 gated as Live CD45 + Lineage negative (CD3 − CD5 − NK1.1 − B220 − CD11b − CD11c − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + cells. Data are shown as individual values or mean ± SEM. Statistical tests performed: (A, B, C, and E) one-way ANOVA with multiple comparisons, (G) Mann–Whitney t test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were stained with the following cell surface antibodies and using the conjugates indicated in the figure labels and utilized for analysis with a BD Fortessa or cell-sorting with a BD Aria Influx; CD127 (IL-7Rα, Brilliant Violet 421, PE, or FITC, clone A7R34; eBioscience), ST2 (IL-33R Biotin; clone RMST2-33; eBioscience), CD45 (brilliant violet 650; clone 30-F11; BioLegend), CD3 (PerCP-Cyanine 5.5 or PE/Cy7; clone 145-2C11), CD5 (PerCP-Cyanine 5.5 or PE/Cy7, clone 53-7.3; BioLegend) NK1.1 (PerPC- Cyanine 5.5, BV395, or PE/Cy7, clone PK136; eBioscience), CD90-2 (Alexa Fluor 700 AM; clone 30-H12; BioLegend), B220 (CD45R, APC-e Fluor 780, clone RA3-6B2; eBioscience), CD11b (super bright 600, APC-e Fluor 780; clone M1/70; Invitrogen), CD11c (APC-e Fluor 780, clone N418; eBioscience), CD4 (super bright 600, clone RM4-5; eBioscience; BV395, clone GK1.5; eBioscience), SA-APC (streptavidin APC, eBioscience), SA-SB600 (streptavidin super bright 600; eBioscience), CD98 (Alexa Fluor 647; clone RL388; BioLegend), CD8α (FITC; clone 53-6.7; BioLegend), KLRG1 (PeCyanine 7, Invitrogen, FITC, or Pe-eFlour 610, clone 2F1; eBioscience), and MHCII (eFluor 450; clone M5/114,15.2; Invitrogen).

Techniques: Infection, MANN-WHITNEY

ILC2 are preferentially poised to import large neutral amino acids. (A) Analysis of the intracellular amino acid content of ILC2 sort-purified from IL-33–treated mice ( n = 3 independent replicates of cells pooled from two mice and representative of two independent experiments). (B) Comparison of mean expression of amino acid transporter-associated genes in ILC2 and CCR6 + ILC3 (ILC3) from public data ( http://www.immgen.org ). (C and D) (C) Representative gating and (D) surface expression of KLRG1 and CD98 on CD4 + T cells (black), ILC2 (red), and ILC3 (green) from small intestinal lamina propria (siLPL). (E) Representative flow plots demonstrating co-expression of GATA-3 and CD98 in lung and siLPL amongst total CD45 + cells. (F) Expression of CD98 on CD4 + T cells (black), B220 + B cells (white), ILC3 (green), and ILC2 (red) in siLPL, colon lamina propria (cLPL), mLN, lung, white adipose tissue (fat), and meninges. (C–F) n = 3 and representative of at least three independent experiments. (G) Relative expression of Slc7a5 and Slc7a8 in small intestinal ILC subsets and B cells, normalized to CCR6 + ILC3 ( n = 4 per group and representative of at least two independent experiments). (H) Representative histogram of Kynurenine uptake in lung ILC2 incubated for 5 min with either HBSS alone (negative control), 200 μM Kynurenine (Kyn) or Kynurenine plus 10 mM BCH. (I) Kynurenine uptake in ILC2 from naive (Ctrl) or IL-33–treated mice or incubated with Kynurenine and BCH ( n = 3 per group and representative of two independent experiments). (J) Kynurenine uptake in lung ILC2 in the presence or absence of excess (5 mM) Lysine (Lys), Leucine (Leu), or 10 mM BCH ( n = 3 replicates per condition, and representative of at least three independent experiments). ILC2 gated as in C or for G–J as Live CD45 + Lineage negative (CD3 − CD5 − NK1.1 − B220 − CD11b − CD11c − ) CD127 + CD90.2 + ST2 + KLRG1 + cells.

Journal: The Journal of Experimental Medicine

Article Title: Amino acid availability acts as a metabolic rheostat to determine the magnitude of ILC2 responses

doi: 10.1084/jem.20221073

Figure Lengend Snippet: ILC2 are preferentially poised to import large neutral amino acids. (A) Analysis of the intracellular amino acid content of ILC2 sort-purified from IL-33–treated mice ( n = 3 independent replicates of cells pooled from two mice and representative of two independent experiments). (B) Comparison of mean expression of amino acid transporter-associated genes in ILC2 and CCR6 + ILC3 (ILC3) from public data ( http://www.immgen.org ). (C and D) (C) Representative gating and (D) surface expression of KLRG1 and CD98 on CD4 + T cells (black), ILC2 (red), and ILC3 (green) from small intestinal lamina propria (siLPL). (E) Representative flow plots demonstrating co-expression of GATA-3 and CD98 in lung and siLPL amongst total CD45 + cells. (F) Expression of CD98 on CD4 + T cells (black), B220 + B cells (white), ILC3 (green), and ILC2 (red) in siLPL, colon lamina propria (cLPL), mLN, lung, white adipose tissue (fat), and meninges. (C–F) n = 3 and representative of at least three independent experiments. (G) Relative expression of Slc7a5 and Slc7a8 in small intestinal ILC subsets and B cells, normalized to CCR6 + ILC3 ( n = 4 per group and representative of at least two independent experiments). (H) Representative histogram of Kynurenine uptake in lung ILC2 incubated for 5 min with either HBSS alone (negative control), 200 μM Kynurenine (Kyn) or Kynurenine plus 10 mM BCH. (I) Kynurenine uptake in ILC2 from naive (Ctrl) or IL-33–treated mice or incubated with Kynurenine and BCH ( n = 3 per group and representative of two independent experiments). (J) Kynurenine uptake in lung ILC2 in the presence or absence of excess (5 mM) Lysine (Lys), Leucine (Leu), or 10 mM BCH ( n = 3 replicates per condition, and representative of at least three independent experiments). ILC2 gated as in C or for G–J as Live CD45 + Lineage negative (CD3 − CD5 − NK1.1 − B220 − CD11b − CD11c − ) CD127 + CD90.2 + ST2 + KLRG1 + cells.

Article Snippet: Cells were stained with the following cell surface antibodies and using the conjugates indicated in the figure labels and utilized for analysis with a BD Fortessa or cell-sorting with a BD Aria Influx; CD127 (IL-7Rα, Brilliant Violet 421, PE, or FITC, clone A7R34; eBioscience), ST2 (IL-33R Biotin; clone RMST2-33; eBioscience), CD45 (brilliant violet 650; clone 30-F11; BioLegend), CD3 (PerCP-Cyanine 5.5 or PE/Cy7; clone 145-2C11), CD5 (PerCP-Cyanine 5.5 or PE/Cy7, clone 53-7.3; BioLegend) NK1.1 (PerPC- Cyanine 5.5, BV395, or PE/Cy7, clone PK136; eBioscience), CD90-2 (Alexa Fluor 700 AM; clone 30-H12; BioLegend), B220 (CD45R, APC-e Fluor 780, clone RA3-6B2; eBioscience), CD11b (super bright 600, APC-e Fluor 780; clone M1/70; Invitrogen), CD11c (APC-e Fluor 780, clone N418; eBioscience), CD4 (super bright 600, clone RM4-5; eBioscience; BV395, clone GK1.5; eBioscience), SA-APC (streptavidin APC, eBioscience), SA-SB600 (streptavidin super bright 600; eBioscience), CD98 (Alexa Fluor 647; clone RL388; BioLegend), CD8α (FITC; clone 53-6.7; BioLegend), KLRG1 (PeCyanine 7, Invitrogen, FITC, or Pe-eFlour 610, clone 2F1; eBioscience), and MHCII (eFluor 450; clone M5/114,15.2; Invitrogen).

Techniques: Purification, Expressing, Incubation, Negative Control

Amino acid transport in ILC2 and other tissue resident lymphocytes. (A–D) CD98 MFI in tissue-resident lymphocytes isolated from naive mice tissues. (A) Lung, (B) small intestinal lamina propria (siLPL), (C) small intestinal intraepithelial lymphocytes (siIEL). (D) Representative flow plot comparing ILC2 to ILC3 and tissue-resident T cell subsets in the siLPL. Cells gated as Live CD45 + and further subgated as: Lineage negative (CD3 − CD5 − NK1.1 − B220 − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + - ILC2; Lineage negative (CD3 − CD5 − NK1.1 − B220 − ) CD127 + CD90.2 + GATA-3 lo RORγt + - ILC3; CD3 + CD5 + Tcrβ + CD4 + - total CD4 + T cells; CD3 + CD5 + CD4 + RORγt + - Th17; CD3 + CD5 + CD4 + - FoxP3 + Treg; CD3 + CD5 + Tcrβ + CD8 + - CD8 + T cells; CD3 + CD5 + Tcrγδ + - gd T cells; CD3 + CD5 + Tcrγδ − CD27 ± CD1d (PBS-57)-Tet + - natural killer T (NKT) cells; CD3 − CD5 − B220 − NK1.1 + - NK cells. (A–D) n = 4 mice per group and representative of two independent experiments. (E and F) Enrichment of ILCs by CD98 + GATA-3 + gating from total CD45 + lymphocytes ( n = 4–5 mice per group, representative of at least three independent experiments). (G) CD98 expression on CD127 + CD25 + ST2 + ILC2 progenitors (ILC2P; red) and Lineage positive bone marrow cells (Lin + ; gray). n = 4 mice per group and representative of two independent experiments. (H) Representative histogram of Kynurenine uptake in small intestinal or mLN ILC2 incubated for 5 min with 200 μM Kynurenine (Kyn) or Kynurenine plus 10 mM BCH. (I) Quantification of n = 3 per group, representative of two independent experiments. (J) Exemplar CD98 expression and Kynurenine uptake in CD4 + T cells from the mLN cultured with either rIL-2 and rIL-7 alone, anti-CD3 and anti-CD28 monoclonal antibodies, or anti-CD3 and anti-CD28 plus BCH ( n = 2–4 replicates per group, representative of two independent experiments). (K) Kynurenine uptake in lung ILC2 in the presence or absence of excess Leucine (Leu), Alanine (Ala), or Methionine (Met). n = 3–4 replicates per condition, and representative of two independent experiments.

Journal: The Journal of Experimental Medicine

Article Title: Amino acid availability acts as a metabolic rheostat to determine the magnitude of ILC2 responses

doi: 10.1084/jem.20221073

Figure Lengend Snippet: Amino acid transport in ILC2 and other tissue resident lymphocytes. (A–D) CD98 MFI in tissue-resident lymphocytes isolated from naive mice tissues. (A) Lung, (B) small intestinal lamina propria (siLPL), (C) small intestinal intraepithelial lymphocytes (siIEL). (D) Representative flow plot comparing ILC2 to ILC3 and tissue-resident T cell subsets in the siLPL. Cells gated as Live CD45 + and further subgated as: Lineage negative (CD3 − CD5 − NK1.1 − B220 − ) CD127 + CD90.2 + GATA-3 hi KLRG1 + - ILC2; Lineage negative (CD3 − CD5 − NK1.1 − B220 − ) CD127 + CD90.2 + GATA-3 lo RORγt + - ILC3; CD3 + CD5 + Tcrβ + CD4 + - total CD4 + T cells; CD3 + CD5 + CD4 + RORγt + - Th17; CD3 + CD5 + CD4 + - FoxP3 + Treg; CD3 + CD5 + Tcrβ + CD8 + - CD8 + T cells; CD3 + CD5 + Tcrγδ + - gd T cells; CD3 + CD5 + Tcrγδ − CD27 ± CD1d (PBS-57)-Tet + - natural killer T (NKT) cells; CD3 − CD5 − B220 − NK1.1 + - NK cells. (A–D) n = 4 mice per group and representative of two independent experiments. (E and F) Enrichment of ILCs by CD98 + GATA-3 + gating from total CD45 + lymphocytes ( n = 4–5 mice per group, representative of at least three independent experiments). (G) CD98 expression on CD127 + CD25 + ST2 + ILC2 progenitors (ILC2P; red) and Lineage positive bone marrow cells (Lin + ; gray). n = 4 mice per group and representative of two independent experiments. (H) Representative histogram of Kynurenine uptake in small intestinal or mLN ILC2 incubated for 5 min with 200 μM Kynurenine (Kyn) or Kynurenine plus 10 mM BCH. (I) Quantification of n = 3 per group, representative of two independent experiments. (J) Exemplar CD98 expression and Kynurenine uptake in CD4 + T cells from the mLN cultured with either rIL-2 and rIL-7 alone, anti-CD3 and anti-CD28 monoclonal antibodies, or anti-CD3 and anti-CD28 plus BCH ( n = 2–4 replicates per group, representative of two independent experiments). (K) Kynurenine uptake in lung ILC2 in the presence or absence of excess Leucine (Leu), Alanine (Ala), or Methionine (Met). n = 3–4 replicates per condition, and representative of two independent experiments.

Article Snippet: Cells were stained with the following cell surface antibodies and using the conjugates indicated in the figure labels and utilized for analysis with a BD Fortessa or cell-sorting with a BD Aria Influx; CD127 (IL-7Rα, Brilliant Violet 421, PE, or FITC, clone A7R34; eBioscience), ST2 (IL-33R Biotin; clone RMST2-33; eBioscience), CD45 (brilliant violet 650; clone 30-F11; BioLegend), CD3 (PerCP-Cyanine 5.5 or PE/Cy7; clone 145-2C11), CD5 (PerCP-Cyanine 5.5 or PE/Cy7, clone 53-7.3; BioLegend) NK1.1 (PerPC- Cyanine 5.5, BV395, or PE/Cy7, clone PK136; eBioscience), CD90-2 (Alexa Fluor 700 AM; clone 30-H12; BioLegend), B220 (CD45R, APC-e Fluor 780, clone RA3-6B2; eBioscience), CD11b (super bright 600, APC-e Fluor 780; clone M1/70; Invitrogen), CD11c (APC-e Fluor 780, clone N418; eBioscience), CD4 (super bright 600, clone RM4-5; eBioscience; BV395, clone GK1.5; eBioscience), SA-APC (streptavidin APC, eBioscience), SA-SB600 (streptavidin super bright 600; eBioscience), CD98 (Alexa Fluor 647; clone RL388; BioLegend), CD8α (FITC; clone 53-6.7; BioLegend), KLRG1 (PeCyanine 7, Invitrogen, FITC, or Pe-eFlour 610, clone 2F1; eBioscience), and MHCII (eFluor 450; clone M5/114,15.2; Invitrogen).

Techniques: Isolation, Expressing, Incubation, Cell Culture

Compound deletion of Slc7a5 and Slc7a8 reduces ILC2 effector responses. (A and B) (A) Representative expression of CD98 and (B) quantification of CD98 MFI on lung ILC2 from IL-33–treated Red5 Cre controls, Red5 x Slc7a5 fl/fl , Red5 x Slc7a8 fl/fl , or Red5 x Slc7a5 fl/fl x Slc7a8 fl/fl (DBL f/f ). (C–E) (C) Representative flow cytometry plots and (D) quantification of Ki-67 + ILC2, and (E) ILC2 numbers in control and DBL f/f mice. (F and G) (F) Representative flow cytometry plots and (G) quantification of IL-5+ IL-13 + ILC2 in lung ILC2. (H) Lung Live CD45 + CD3 − CD5 − NK1.1 − B220 − Ly6C − Ly6G − CD11c − Siglec F + Eosinophil numbers in control and DBL f/f mice. (A–H) All data representative of pooled data of n = 6–8 mice per group from at least two independent experiments. (I and J) (I) Representative flow cytometry plots, and (J) quantification of IL-13 expression of naive and IL-33 activated lung ILC2 in the presence or absence of BCH. Data are representative of two independent experiments with n = 3 replicates per culture condition. ILC2 gated as in . Data shown as individual values or mean ± SEM. Statistical tests performed: (B, D, and G) one-way ANOVA with multiple comparisons; (E and H) unpaired t test. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Journal: The Journal of Experimental Medicine

Article Title: Amino acid availability acts as a metabolic rheostat to determine the magnitude of ILC2 responses

doi: 10.1084/jem.20221073

Figure Lengend Snippet: Compound deletion of Slc7a5 and Slc7a8 reduces ILC2 effector responses. (A and B) (A) Representative expression of CD98 and (B) quantification of CD98 MFI on lung ILC2 from IL-33–treated Red5 Cre controls, Red5 x Slc7a5 fl/fl , Red5 x Slc7a8 fl/fl , or Red5 x Slc7a5 fl/fl x Slc7a8 fl/fl (DBL f/f ). (C–E) (C) Representative flow cytometry plots and (D) quantification of Ki-67 + ILC2, and (E) ILC2 numbers in control and DBL f/f mice. (F and G) (F) Representative flow cytometry plots and (G) quantification of IL-5+ IL-13 + ILC2 in lung ILC2. (H) Lung Live CD45 + CD3 − CD5 − NK1.1 − B220 − Ly6C − Ly6G − CD11c − Siglec F + Eosinophil numbers in control and DBL f/f mice. (A–H) All data representative of pooled data of n = 6–8 mice per group from at least two independent experiments. (I and J) (I) Representative flow cytometry plots, and (J) quantification of IL-13 expression of naive and IL-33 activated lung ILC2 in the presence or absence of BCH. Data are representative of two independent experiments with n = 3 replicates per culture condition. ILC2 gated as in . Data shown as individual values or mean ± SEM. Statistical tests performed: (B, D, and G) one-way ANOVA with multiple comparisons; (E and H) unpaired t test. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Cells were stained with the following cell surface antibodies and using the conjugates indicated in the figure labels and utilized for analysis with a BD Fortessa or cell-sorting with a BD Aria Influx; CD127 (IL-7Rα, Brilliant Violet 421, PE, or FITC, clone A7R34; eBioscience), ST2 (IL-33R Biotin; clone RMST2-33; eBioscience), CD45 (brilliant violet 650; clone 30-F11; BioLegend), CD3 (PerCP-Cyanine 5.5 or PE/Cy7; clone 145-2C11), CD5 (PerCP-Cyanine 5.5 or PE/Cy7, clone 53-7.3; BioLegend) NK1.1 (PerPC- Cyanine 5.5, BV395, or PE/Cy7, clone PK136; eBioscience), CD90-2 (Alexa Fluor 700 AM; clone 30-H12; BioLegend), B220 (CD45R, APC-e Fluor 780, clone RA3-6B2; eBioscience), CD11b (super bright 600, APC-e Fluor 780; clone M1/70; Invitrogen), CD11c (APC-e Fluor 780, clone N418; eBioscience), CD4 (super bright 600, clone RM4-5; eBioscience; BV395, clone GK1.5; eBioscience), SA-APC (streptavidin APC, eBioscience), SA-SB600 (streptavidin super bright 600; eBioscience), CD98 (Alexa Fluor 647; clone RL388; BioLegend), CD8α (FITC; clone 53-6.7; BioLegend), KLRG1 (PeCyanine 7, Invitrogen, FITC, or Pe-eFlour 610, clone 2F1; eBioscience), and MHCII (eFluor 450; clone M5/114,15.2; Invitrogen).

Techniques: Expressing, Flow Cytometry