Journal: The Journal of Neuroscience

Article Title: Somatotopy of Mouse Spinothalamic Innervation and the Localization of a Noxious Stimulus Requires Deleted in Colorectal Carcinoma Expression by Phox2a Neurons

doi: 10.1523/JNEUROSCI.1164-22.2022

Figure Lengend Snippet: Resources and reagents used

Article Snippet: Reagent or resource Source/reference Identifier Mice (MGI notation) Dcc flox Krimpenfort et al. 2012 MGI: 3665466 Phox2a Cre Roome et al. 2022 RRID: NA Ai14 B6;129S6-Gt (ROSA)26Sortm14(CAG- tdTomato ) Hze/J The Jackson Laboratory Catalog #JAX:007908 RRID: IMSR_JAX:007908 Cdx2 FlpO Tg(CDX2-flpo)#Gld Dr. Martyn Goulding, Salk Institute, San Diego MGI: 5911680 RRID: NA Ai65 B6;129S-Gt ROSA)26Sortm65.1(CAG- dTomato) Hze/J The Jackson Laboratory Catalog #JAX:021875 RRID: IMSR_JAX:021875 Ntn1 Bgeo Skarnes et al., 1995 RRID: NA 129S1/SvImJ The Jackson Laboratory Catalog #JAX:002448 RRID: IMSR_JAX:002448 B6C3F1/J The Jackson Laboratory Catalog #JAX: 100010 RRID: IMSR_JAX:100010 Primary antibodies Goat anti-mouse DCC (1:500) R&D Systems Catalog #AF844 RRID: AB_2089765 Rabbit anti-Phox2a (1:10,000 from frozen stock) Jean-François Brunet (École Normale Supérieure, Paris) ( Tiveron et al., 1996 ) RRID: AB_2315159 Rabbit anti-RFP (red fluorescent protein; 1:1000) Rockland Catalog #600-401-379 RRID: AB_2209751 Goat anti-tdT (red fluorescent protein; 1:300) SICGEN Catalog #AB8181-200 RRID: AB_2722750 Rabbit anti-GFP (1:1000) Life Technologies Catalog #A-11122 RRID: AB_221569 Goat anti-netrin-1 (1:200) R&D Systems Catalog #AF1109, RRID: AB_2298775 Rabbit anti-c- fos (1:500) Cell Signaling Technology Catalog #2250S RRID: AB_2247211 Rabbit anti-Pax6 (1:500) Millipore Catalog #AB2237, RRID: AB_1587367 Rabbit anti-Sox2 (1:200) Abcam Catalog #97959 RRID: AB_2341193 Secondary antibodies Alexa 488 Donkey anti-Rabbit (1:500) Jackson ImmunoResearch Laboratories Catalog #711–545-152 Lot #141848 RRID: AB_2313584 Alexa 488 Donkey anti-Goat (1:500) Jackson ImmunoResearch Laboratories Catalog #705–545-147 Lot #136089 RRID: AB_2336933 Cy3 Donkey anti-Rabbit (1:500) Jackson ImmunoResearch Laboratories Catalog #711–165-152 Lot #138270 RRID: AB_2307443 Cy3 Donkey anti-Goat (1:500) Jackson ImmunoResearch Laboratories Catalog #705–165-147 Lot #134527 RRID: AB_2307351 Cy5 Donkey anti-Rabbit (1:500) Jackson ImmunoResearch Laboratories Catalog #711–175-152 Lot #138336 RRID: AB_2340607 Cy5 Donkey anti-Goat (1:500) Jackson ImmunoResearch Laboratories Catalog #: 705–175-147 Lot#: 134531 RRID: AB_2340415 Chemicals ( E )-Capsaicin Tocris Bioscience Catalog #0462 Lot #7A/218361 RRID: NA 4′,6-diamidino-2-phenylindole (DAPI) Thermo Fisher Scientific Catalog #D1306 RRID: NA Alexa 488-conjugated Choleratoxin B Thermo Fisher Scientific Catalog # C22841 Lot #2038245 RRID: NA X-gal Millipore Sigma Catalog #7240-90-6 RRID: NA MOWIOL Millipore Sigma Catalog #81381 RRID: NA Open in a separate window MGI, Mouse Genome Informatics; NA, Not applicable.

Techniques:

Journal: Cell reports

Article Title: Innate Lymphoid Cells Play a Pathogenic Role in Pericarditis

doi: 10.1016/j.celrep.2020.02.040

Figure Lengend Snippet: (A) Representative images of H&E-stained heart sections of the median mice treated with either PBS or IL-33. Areas marked by rectangles are shown as enlarged images in the right panels. Bars: 1 mm (left) and 100 μm (right). (B) Representative M-mode pictures of animals treated with PBS or IL-33. Bars: blue, 1 mm; white, 0.1 s. LVESD, left ventricular end-systolic diameter; LVEDD, left ventricular end-diastolic diameter. (C and D) Ejection fraction (EF) (C) and fractional shortening (FS) (D) of WT mice treated with PBS or IL-33. (E and F) Isovolumetric relaxation time (IVRT) (E) and myocardial performance indexes (MPI) (F) of the heart from mice treated with PBS or IL-33 were assessed by Doppler echocardiography on day 9 post-PBS or IL-33 treatment. (G) Total number of heart-infiltrating CD45 + leukocytes was determined by flow cytometry. (H–J) Number of (H) ILC2s, (I) eosinophils, and (J) mast cells in the hearts. Absolute cell counts per heart were calculated using counting beads for flow cytometry (see ). (K) Image of the right ventricle of the heart from Rag2 −/− mice treated with IL-33. Green and red dots represent KLRG1 + ILC2s (white arrows) and SiglecF + eosinophils (red arrowheads), respectively. Scale bar: 500 μm. Data are representative of three independent experiments. Data are displayed as the means. Unpaired t test (C–J) was used for statistical analysis. *p < 0.05; **p < 0.01; ***p < 0.001. See also and and .

Article Snippet: Primary and secondary antibody staining were performed using 10 μg/ml anti-mouse SiglecF (BD) and KLRG1 (ProSci) antibodies and 20 μg/ml anti-rat and anti-hamster IgG conjugated with fluorophores (Thermo Fisher Scientific), respectively, in Visikol HISTO antibody buffer (Visikol) according to manufacturer’s protocols.

Techniques: Staining, Flow Cytometry

Journal: Cell reports

Article Title: Innate Lymphoid Cells Play a Pathogenic Role in Pericarditis

doi: 10.1016/j.celrep.2020.02.040

Figure Lengend Snippet: (A) Representative images of H&E-stained heart sections of WT (left), Rag2 −/− (center), and Rag2 −/− Il2rg −/− (right) mice. Bars: 100 μm. (B) Severity of pericarditis was scored on H&E-stained heart sections. (C) Total number of heart-infiltrating CD45 + leukocytes was determined by flow cytometry. (D and E) Number of (D) ILC2s and (E) eosinophils in the hearts. (F) Representative flow cytometry plots of ILC2s found in the hearts of naive Rag2 −/− Il2rg −/− mice and Rag2 −/− Il2rg −/− mice injected with media or ILC2s followed by IL-33 treatment. (G) Number of heart-infiltrating CD45.2 + leukocytes. (H) Representative flow cytometry plots of CD45.2 + cells. Gates show the frequency of CD11b + SiglecF + eosinophils in the hearts of naive Rag2 −/− Il2rg −/− mice and Rag2 −/− Il2rg −/− mice injected with media or ILC2s followed by IL-33 treatment. (I) Number of eosinophils in the hearts. Both male and female mice were used in each group in (B). Data are representative of two independent experiments and displayed as the mean. Kruskal-Wallis H test (B) or one-way ANOVA followed by Tukey’s post hoc test (C–E, G, and I) was used for statistical analysis. *p < 0.05; **p < 0.01; ***p < 0.001. See also .

Article Snippet: Primary and secondary antibody staining were performed using 10 μg/ml anti-mouse SiglecF (BD) and KLRG1 (ProSci) antibodies and 20 μg/ml anti-rat and anti-hamster IgG conjugated with fluorophores (Thermo Fisher Scientific), respectively, in Visikol HISTO antibody buffer (Visikol) according to manufacturer’s protocols.

Techniques: Staining, Flow Cytometry, Injection

Journal: Cell reports

Article Title: Innate Lymphoid Cells Play a Pathogenic Role in Pericarditis

doi: 10.1016/j.celrep.2020.02.040

Figure Lengend Snippet: (A) Representative flow cytometry plot of CD45 + CD11b + cells. Gates show CD11b + SiglecF + eosinophils and CD11b + Ly6G + neutrophils in the heart, mediastinal cavity, and blood of WT naive mice. (B) Frequency of eosinophils in the heart, mediastinal cavity, and blood of WT naive mice. (C) Representative flow cytometry plot of CD45 + CD11b + cells. Gates show CD11b + SiglecF + eosinophils and CD11b + Ly6G + neutrophils in the heart, mediastinal cavity, and blood of IL-5Tg naive mice. (D) Frequency of eosinophils in the heart, mediastinal cavity, and blood of IL-5Tg naive mice. (E) Schematic description of eosinophil transfer to eosinophil-deficient ΔdblGATA1 mice. (F) Flow cytometry plots of CTV- or CTFR-labeled eosinophils. (G) Flow cytometry plot of CD45 + cells in the heart. Gate shows CD11b + SiglecF + eosinophils found in the heart of ΔdblGATA1 mice treated with IL-33 after eosinophil transfer in the mediastinal cavity and i.v. (H) CTV- or CTFR-labeled CD11b + SiglecF + eosinophils found in the heart of ΔdblGATA1 mice treated with IL-33 after eosinophil transfer. (I) Frequency of CTV- or CTFR-labeled eosinophils found in the heart of ΔdblGATA1 mice treated with IL-33. Concatenated samples (n = 7) are shown in (G) and (H). Both male and female mice were used in (G)–(I). Data are representative of two independent experiments and displayed as the mean (B and D) or the mean with SD (I). One-way ANOVA followed by Tukey’s post hoc test (B and D) or unpaired t test (I) was used for statistical analysis. **p < 0.01. See also .

Article Snippet: Primary and secondary antibody staining were performed using 10 μg/ml anti-mouse SiglecF (BD) and KLRG1 (ProSci) antibodies and 20 μg/ml anti-rat and anti-hamster IgG conjugated with fluorophores (Thermo Fisher Scientific), respectively, in Visikol HISTO antibody buffer (Visikol) according to manufacturer’s protocols.

Techniques: Flow Cytometry, Labeling

Journal: EMBO Molecular Medicine

Article Title: Combining chemotherapeutic agents and netrin-1 interference potentiates cancer cell death

doi: 10.1002/emmm.201302654

Figure Lengend Snippet: Netrin-1 and its dependence receptors are upregulated upon Doxorubicin treatment. Source data is available for this figure in the Supporting Information.

Article Snippet: Endogenous netrin-1 was stained using rat monoclonal α-netrin-1 antibody (R&D systems) and Alexa-488 Donkey anti-rat IgG (Molecular probes). p53 protein was stained using the mouse monoclonal α-p53-DO-1 antibody.

Techniques:

Journal: EMBO Molecular Medicine

Article Title: Combining chemotherapeutic agents and netrin-1 interference potentiates cancer cell death

doi: 10.1002/emmm.201302654

Figure Lengend Snippet: Netrin-1 and its receptors expression is increased in several cancer cell lines and in ovarian tumours upon treatment with cytotoxic drugs.

Article Snippet: Endogenous netrin-1 was stained using rat monoclonal α-netrin-1 antibody (R&D systems) and Alexa-488 Donkey anti-rat IgG (Molecular probes). p53 protein was stained using the mouse monoclonal α-p53-DO-1 antibody.

Techniques: Expressing

Journal: EMBO Molecular Medicine

Article Title: Combining chemotherapeutic agents and netrin-1 interference potentiates cancer cell death

doi: 10.1002/emmm.201302654

Figure Lengend Snippet: Netrin-1 silencing sensitizes A549R cells to Doxorubicin and induces apoptotic cell death via UNC5B receptor.

Article Snippet: Endogenous netrin-1 was stained using rat monoclonal α-netrin-1 antibody (R&D systems) and Alexa-488 Donkey anti-rat IgG (Molecular probes). p53 protein was stained using the mouse monoclonal α-p53-DO-1 antibody.

Techniques:

Journal: EMBO Molecular Medicine

Article Title: Combining chemotherapeutic agents and netrin-1 interference potentiates cancer cell death

doi: 10.1002/emmm.201302654

Figure Lengend Snippet: Interference to netrin-1 and its receptors interaction sensitizes tumour cells to cytotoxic drugs.

Article Snippet: Endogenous netrin-1 was stained using rat monoclonal α-netrin-1 antibody (R&D systems) and Alexa-488 Donkey anti-rat IgG (Molecular probes). p53 protein was stained using the mouse monoclonal α-p53-DO-1 antibody.

Techniques:

Journal: EMBO Molecular Medicine

Article Title: Combining chemotherapeutic agents and netrin-1 interference potentiates cancer cell death

doi: 10.1002/emmm.201302654

Figure Lengend Snippet: Tumour growth inhibiting effect of combining netrin-1 interference and Doxorubicin.

Article Snippet: Endogenous netrin-1 was stained using rat monoclonal α-netrin-1 antibody (R&D systems) and Alexa-488 Donkey anti-rat IgG (Molecular probes). p53 protein was stained using the mouse monoclonal α-p53-DO-1 antibody.

Techniques:

Journal: EMBO Molecular Medicine

Article Title: Combining chemotherapeutic agents and netrin-1 interference potentiates cancer cell death

doi: 10.1002/emmm.201302654

Figure Lengend Snippet: Netrin-1 upregulation is p53-dependent.

Article Snippet: Endogenous netrin-1 was stained using rat monoclonal α-netrin-1 antibody (R&D systems) and Alexa-488 Donkey anti-rat IgG (Molecular probes). p53 protein was stained using the mouse monoclonal α-p53-DO-1 antibody.

Techniques: