goat anti mouse  (Santa Cruz Biotechnology)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Name:
    goat anti mouse IgG B
    Description:
    supplied at 200 µg in 0 5 ml volume pre adsorbed biotin conjugated secondary antibody recommended for use in immunohistochemical staining recommended dilution range 1 100 1 400
    Catalog Number:
    SC-2039
    Price:
    None
    Category:
    Antibodies Conventional Secondary IgGs goat anti mouse IgG B
    Buy from Supplier


    Structured Review

    Santa Cruz Biotechnology goat anti mouse
    supplied at 200 µg in 0 5 ml volume pre adsorbed biotin conjugated secondary antibody recommended for use in immunohistochemical staining recommended dilution range 1 100 1 400
    https://www.bioz.com/result/goat anti mouse/product/Santa Cruz Biotechnology
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse - by Bioz Stars, 2021-04
    97/100 stars

    Images

    Related Articles

    Immunoprecipitation:

    Article Title: Aldosterone and vasopressin affect ?- and ?-ENaC mRNA translation
    Article Snippet: Briefly, mCCD cells were stimulated as described, followed by lysis in a buffer containing 10 mM HEPES, pH 7.9, 1.5 mM MgCl2, 10 mM KCl, 0.5 mM DTT, 0.1% Nonidet P-40, 50 mM NaF, 10 mM Na3 VO4 , 10 mM sodium pyrophosphate, 50 mM disodium glycerol phosphate and 100 U/ml RNasin. .. Subsequently, cell lysates were immunoprecipitated with 2 μg of either a monoclonal anti-HuR antibody (Santa Cruz) or with the same amount of mouse IgG (Santa Cruz) as negative control overnight at 4°C. .. To normalize for equal input of RNA before subsequent purification steps, the same amount of extract was subjected to total RNA isolation as described.

    Negative Control:

    Article Title: Aldosterone and vasopressin affect ?- and ?-ENaC mRNA translation
    Article Snippet: Briefly, mCCD cells were stimulated as described, followed by lysis in a buffer containing 10 mM HEPES, pH 7.9, 1.5 mM MgCl2, 10 mM KCl, 0.5 mM DTT, 0.1% Nonidet P-40, 50 mM NaF, 10 mM Na3 VO4 , 10 mM sodium pyrophosphate, 50 mM disodium glycerol phosphate and 100 U/ml RNasin. .. Subsequently, cell lysates were immunoprecipitated with 2 μg of either a monoclonal anti-HuR antibody (Santa Cruz) or with the same amount of mouse IgG (Santa Cruz) as negative control overnight at 4°C. .. To normalize for equal input of RNA before subsequent purification steps, the same amount of extract was subjected to total RNA isolation as described.

    Article Title: TLR4 Activation Promotes Podocyte Injury and Interstitial Fibrosis in Diabetic Nephropathy
    Article Snippet: .. Sections were then incubated with 10% normal horse serum followed by 60-minute incubation with primary antibodies: a rat anti-mouse CD68 antibody (ABD Serotec Inc., Oxford, UK), a rat anti-mouse F4/80 (ABD Serotec Inc.), rabbit anti-mouse-TLR4 antibody (Invitrogen, Catalogue No. 48-2300) for glomerular , goat anti-mouse-TLR4 antibody (Santa Cruz Biotechnology, Catalogue No sc-12511) for tubular , rabbit anti-WT1 antibody (Abcam, Cambridge, UK), or concentration-matched IgG as an isotype negative control. .. The sections were exposed to H2 O2 and then incubated with biotinylated anti-rat IgG or anti-rabbit IgG (BD Biosciences, Pharmingen), or anti-goat IgG (Vector Laboratories Inc).

    other:

    Article Title: Loss of CLN7 results in depletion of soluble lysosomal proteins and impaired mTOR reactivation
    Article Snippet: Primary antibodies were: goat anti-mouse cathepsin B (Neuromics, GT15047), goat anti-mouse cathepsin D (Santa Cruz, sc6486), monoclonal mouse anti-α-tubulin (Sigma, T9026), polyclonal rabbit anti-Cln5 (Abcam, ab170899), polyclonal goat anti-cathepsin S (Santa-Cruz, sc6503), polyclonal goat anti-mouse Dpp7 (R & D, AF3436), polyclonal rabbit anti-glucocerebrosidase (Sigma, G4171), polyclonal rabbit anti-LC3B/MAP1LC3B (Novusbio, NB100–2220), polyclonal rabbit anti-p62 (Enzo Life Sciences, BML-PW9860), polyclonal rabbit anti-human PPT1 (Millipore, ABS1118), polyclonal rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (Gapdh, Santa Cruz, sc25778), polyclonal rabbit anti-SIDT2 (Thermo Fischer Scientific, PA5–34493).

    Incubation:

    Article Title: TLR4 Activation Promotes Podocyte Injury and Interstitial Fibrosis in Diabetic Nephropathy
    Article Snippet: .. Sections were then incubated with 10% normal horse serum followed by 60-minute incubation with primary antibodies: a rat anti-mouse CD68 antibody (ABD Serotec Inc., Oxford, UK), a rat anti-mouse F4/80 (ABD Serotec Inc.), rabbit anti-mouse-TLR4 antibody (Invitrogen, Catalogue No. 48-2300) for glomerular , goat anti-mouse-TLR4 antibody (Santa Cruz Biotechnology, Catalogue No sc-12511) for tubular , rabbit anti-WT1 antibody (Abcam, Cambridge, UK), or concentration-matched IgG as an isotype negative control. .. The sections were exposed to H2 O2 and then incubated with biotinylated anti-rat IgG or anti-rabbit IgG (BD Biosciences, Pharmingen), or anti-goat IgG (Vector Laboratories Inc).

    Concentration Assay:

    Article Title: TLR4 Activation Promotes Podocyte Injury and Interstitial Fibrosis in Diabetic Nephropathy
    Article Snippet: .. Sections were then incubated with 10% normal horse serum followed by 60-minute incubation with primary antibodies: a rat anti-mouse CD68 antibody (ABD Serotec Inc., Oxford, UK), a rat anti-mouse F4/80 (ABD Serotec Inc.), rabbit anti-mouse-TLR4 antibody (Invitrogen, Catalogue No. 48-2300) for glomerular , goat anti-mouse-TLR4 antibody (Santa Cruz Biotechnology, Catalogue No sc-12511) for tubular , rabbit anti-WT1 antibody (Abcam, Cambridge, UK), or concentration-matched IgG as an isotype negative control. .. The sections were exposed to H2 O2 and then incubated with biotinylated anti-rat IgG or anti-rabbit IgG (BD Biosciences, Pharmingen), or anti-goat IgG (Vector Laboratories Inc).

    Neutralization:

    Article Title: Hepatitis C virus utilizes VLDLR as a novel entry pathway
    Article Snippet: Rabbit anti-NPC1L1 (Cell Signaling), anti-EGFR (Abcam), and mouse anti-core (Institute of Immunology) were purchased also. .. Anti-mouse IgG (Santa Cruz Biotechnology), anti-CD81 (D-4) (Santa Cruz), anti-ApoE (Autogen Bioclear), anti-HCV E2 (AP33), anti-VLDLR (1H10) (Novus), anti-SR-BI (Novus), anti-LDLR (R & D), and anti-NPC1L1 (Santa Cruz) were used for analysis of antibody neutralization. .. Mouse anti-FLAG M2 and chlorpromazine were purchased from Sigma.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    Santa Cruz Biotechnology goat anti fos
    Additional information and analyses for CRISPR-mediated disruption of leptin receptors (LepRs) in AgRP neurons a , sgRNA design for targeting the mouse Lepr genomic locus. b–h , Schematic diagram of AAV pU6-sgRNA Lepr ::pEF1α-FLEX-mCherry (AAV-sgLepR) injected unilaterally into the ARC of <t>Agrp-IRES-Cre::LSL-Cas9-GFP</t> mice ( b ), representative images ( c ) and quantification ( d ) of mCherry and leptin-induced phosphorylated STAT3 (Tyr 705 , pSTAT3) co-immunostaining in the hypothalamus, cell counting of GFP + cells from the ARC suggesting that deletion of Lepr in AgRP neurons does not induce cell death ( e ), representative images of mCherry immunostaining (left) and RNA-ISH against Lepr mRNA (right) indicating efficient deletion of Lepr ( f ), representative images ( g ) and quantification ( h ) of <t>mCherry::Fos</t> co-immunostaining in the ARC of ad libitum fed animals indicating disinhibition of AgRP neurons ( n =3 mice per group). i, j , Schematic diagram of AAV pU6-sgRNA Lepr ::pEF1α-FLEX-mCherry (AAV-sgLepR) injected bilaterally into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP or Agrp-IRES-Cre mice ( i ), quantitative PCR results of Lepr , Agrp , Npy , and Pomc mRNA expression in the ventromedial hypothalamus of fed mice ( j, n=4 mice per group). k, Serum insulin levels in ad libitum fed Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP mice (Cas9 + ) bilaterally injected with AAV-sgLepR virus ( n =4 mice per group). l, CRISPR-mediated deletion of Lepr in AgRP neurons also induces severe obesity in female mice ( n =6 mice per group). m , Representative near-infrared thermal images and quantification of interscapular brown adipose tissue (iBAT) temperature in virus-transduced, ad libitum fed Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP male littermates (Cas9 + ). n , Schematic diagram of assay design to compare mice with global Lepr mutations and mice with specific deletion of Lepr in AgRP neurons. o , p , weekly blood glucose measurement and daily food intake at 8 weeks of age of ad libitum fed, virus-transduced Lepr db/+ ( db/+ ), Lepr db/db ( db/db ), AgRP-IRES-Cre (Cas9 − ), and AgRP-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice ( n =9 mice per group). q-s , Schematic diagram of experiment ( q ), changes in body weight ( r ), and daily food intake ( s) ( 5 days post pump surgery) in STZ-treated, virus-transduced Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice following chronic administration of saline/leptin with osmotic pump ( n =6 mice per group). t–v , Schematic diagram of experiment ( t ), changes of body weight ( u ) and daily food intake ( v ) in non-STZ treated, virus-transduced Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice following 3-day leptin i.p. injection ( n =7 mice per group). Data are mean ± s.e.m. and representative of three independent experiments; * P
    Goat Anti Fos, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti fos/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti fos - by Bioz Stars, 2021-04
    86/100 stars
      Buy from Supplier

    93
    Santa Cruz Biotechnology goat anti mum1 irf 4
    Id + diffuse large B cell lymphomas are infiltrated with Id-specific T cells. (A) Lymphomatous spleen of an Id-Th2→Id + mouse (top) compared with spleen from an Ova-Th2→Id + mouse (bottom). (B) Photo of enlarged mesenteric LN in an Id-Th2→Id + mouse with lymphoma (left). The circled asterisk indicates the normal size of a mesenteric LN in an Ova-Th2→Id + mouse. (right) Liver from Id-Th2→Id + mouse. Arrows indicate tumor nodules. (C) Low and high power magnification (hematoxylin and eosin) of lymphomatous spleen. (D) Lymphoma of the liver with infiltrating CD3 + T cells (brown). (E) Spleen lymphoma were B220 + , <t>IRF-4/MUM1</t> + , and CD138 + (in about half of the mice) and were IgM + IgD + . In the rightmost immunofluorescence picture, DAPI stains the nuclei. (F) Low power magnification of splenic lymphoma, Id + λ2 L chain (red, 2B6 mAb) versus κ (green). (inset) Large Id + lymphoma cell compared with smaller κ + B cell. (G) Id-specific T cells (red, TCR clonotype-specific mAb GB113) and lymphoma cells (green, 2B6 mAb) were often in close contact (yellow). See inset for higher magnification of a synapse.
    Goat Anti Mum1 Irf 4, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mum1 irf 4/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mum1 irf 4 - by Bioz Stars, 2021-04
    93/100 stars
      Buy from Supplier

    93
    Santa Cruz Biotechnology anti cd74
    N terminus and first transmembrane segment of Na v 1.6 are sufficient to direct a reporter protein to the cell surface. Confocal images of HEK293 cells transfected with the reporter constructs and probed with anti-CD4 antibodies are shown. Green , <t>anti-CD74;</t> blue , DAPI. A , extracellular domain of CD74 lacking membrane-targeting N terminus. B , N terminus and first transmembrane segment of Na v 1.6 fused to the extracellular domain of CD74. C , ataxia3 mutation S21P not disrupting surface localization.
    Anti Cd74, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd74/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd74 - by Bioz Stars, 2021-04
    93/100 stars
      Buy from Supplier

    97
    Santa Cruz Biotechnology goat anti mouse igg
    <t>HCt/E-specific</t> antibody production in mice immunized with BoNT/HCt/E DNA vaccine by intradermal electroporation. Mice were immunized 3 times (week 0, 2, 4) with 50 μg of DNA vaccine. Anti-HCt/E <t>IgG</t> levels are expressed as endpoint titers. Each symbol represents data from an individual mouse. The bar represents the mean titer for each group.
    Goat Anti Mouse Igg, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse igg/product/Santa Cruz Biotechnology
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse igg - by Bioz Stars, 2021-04
    97/100 stars
      Buy from Supplier

    Image Search Results


    Additional information and analyses for CRISPR-mediated disruption of leptin receptors (LepRs) in AgRP neurons a , sgRNA design for targeting the mouse Lepr genomic locus. b–h , Schematic diagram of AAV pU6-sgRNA Lepr ::pEF1α-FLEX-mCherry (AAV-sgLepR) injected unilaterally into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP mice ( b ), representative images ( c ) and quantification ( d ) of mCherry and leptin-induced phosphorylated STAT3 (Tyr 705 , pSTAT3) co-immunostaining in the hypothalamus, cell counting of GFP + cells from the ARC suggesting that deletion of Lepr in AgRP neurons does not induce cell death ( e ), representative images of mCherry immunostaining (left) and RNA-ISH against Lepr mRNA (right) indicating efficient deletion of Lepr ( f ), representative images ( g ) and quantification ( h ) of mCherry::Fos co-immunostaining in the ARC of ad libitum fed animals indicating disinhibition of AgRP neurons ( n =3 mice per group). i, j , Schematic diagram of AAV pU6-sgRNA Lepr ::pEF1α-FLEX-mCherry (AAV-sgLepR) injected bilaterally into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP or Agrp-IRES-Cre mice ( i ), quantitative PCR results of Lepr , Agrp , Npy , and Pomc mRNA expression in the ventromedial hypothalamus of fed mice ( j, n=4 mice per group). k, Serum insulin levels in ad libitum fed Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP mice (Cas9 + ) bilaterally injected with AAV-sgLepR virus ( n =4 mice per group). l, CRISPR-mediated deletion of Lepr in AgRP neurons also induces severe obesity in female mice ( n =6 mice per group). m , Representative near-infrared thermal images and quantification of interscapular brown adipose tissue (iBAT) temperature in virus-transduced, ad libitum fed Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP male littermates (Cas9 + ). n , Schematic diagram of assay design to compare mice with global Lepr mutations and mice with specific deletion of Lepr in AgRP neurons. o , p , weekly blood glucose measurement and daily food intake at 8 weeks of age of ad libitum fed, virus-transduced Lepr db/+ ( db/+ ), Lepr db/db ( db/db ), AgRP-IRES-Cre (Cas9 − ), and AgRP-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice ( n =9 mice per group). q-s , Schematic diagram of experiment ( q ), changes in body weight ( r ), and daily food intake ( s) ( 5 days post pump surgery) in STZ-treated, virus-transduced Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice following chronic administration of saline/leptin with osmotic pump ( n =6 mice per group). t–v , Schematic diagram of experiment ( t ), changes of body weight ( u ) and daily food intake ( v ) in non-STZ treated, virus-transduced Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice following 3-day leptin i.p. injection ( n =7 mice per group). Data are mean ± s.e.m. and representative of three independent experiments; * P

    Journal: Nature

    Article Title: Genetic identification of leptin neural circuits in energy and glucose homeostases

    doi: 10.1038/s41586-018-0049-7

    Figure Lengend Snippet: Additional information and analyses for CRISPR-mediated disruption of leptin receptors (LepRs) in AgRP neurons a , sgRNA design for targeting the mouse Lepr genomic locus. b–h , Schematic diagram of AAV pU6-sgRNA Lepr ::pEF1α-FLEX-mCherry (AAV-sgLepR) injected unilaterally into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP mice ( b ), representative images ( c ) and quantification ( d ) of mCherry and leptin-induced phosphorylated STAT3 (Tyr 705 , pSTAT3) co-immunostaining in the hypothalamus, cell counting of GFP + cells from the ARC suggesting that deletion of Lepr in AgRP neurons does not induce cell death ( e ), representative images of mCherry immunostaining (left) and RNA-ISH against Lepr mRNA (right) indicating efficient deletion of Lepr ( f ), representative images ( g ) and quantification ( h ) of mCherry::Fos co-immunostaining in the ARC of ad libitum fed animals indicating disinhibition of AgRP neurons ( n =3 mice per group). i, j , Schematic diagram of AAV pU6-sgRNA Lepr ::pEF1α-FLEX-mCherry (AAV-sgLepR) injected bilaterally into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP or Agrp-IRES-Cre mice ( i ), quantitative PCR results of Lepr , Agrp , Npy , and Pomc mRNA expression in the ventromedial hypothalamus of fed mice ( j, n=4 mice per group). k, Serum insulin levels in ad libitum fed Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP mice (Cas9 + ) bilaterally injected with AAV-sgLepR virus ( n =4 mice per group). l, CRISPR-mediated deletion of Lepr in AgRP neurons also induces severe obesity in female mice ( n =6 mice per group). m , Representative near-infrared thermal images and quantification of interscapular brown adipose tissue (iBAT) temperature in virus-transduced, ad libitum fed Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP male littermates (Cas9 + ). n , Schematic diagram of assay design to compare mice with global Lepr mutations and mice with specific deletion of Lepr in AgRP neurons. o , p , weekly blood glucose measurement and daily food intake at 8 weeks of age of ad libitum fed, virus-transduced Lepr db/+ ( db/+ ), Lepr db/db ( db/db ), AgRP-IRES-Cre (Cas9 − ), and AgRP-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice ( n =9 mice per group). q-s , Schematic diagram of experiment ( q ), changes in body weight ( r ), and daily food intake ( s) ( 5 days post pump surgery) in STZ-treated, virus-transduced Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice following chronic administration of saline/leptin with osmotic pump ( n =6 mice per group). t–v , Schematic diagram of experiment ( t ), changes of body weight ( u ) and daily food intake ( v ) in non-STZ treated, virus-transduced Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice following 3-day leptin i.p. injection ( n =7 mice per group). Data are mean ± s.e.m. and representative of three independent experiments; * P

    Article Snippet: Primary antibodies used in the current study and their dilutions are: rabbit anti-DsRed (Clontech, 1:2000, Cat#632496), chicken anti-mCherry (EnCor Biotechnology, 1:2000, # CPCA-mCherry), chicken anti-GFP (Aves Labs, 1:2000, GFP-1010), rabbit anti-hrGFP (Agilent Technologies, 1:1000, Cat# 240142), goat anti-Fos (Santa Cruz Biotechnology, 1:150, Cat# sc-52-g), rabbit anti-pSTAT3 (Cell Signaling Technology, 1:1000, Cat# 9145S), rabbit phosphor-S6 ribosomal protein (S235/236) (Cell Signaling Technology, 1:1000, Cat# 4858).

    Techniques: CRISPR, Injection, Mouse Assay, Immunostaining, Cell Counting, In Situ Hybridization, Real-time Polymerase Chain Reaction, Expressing

    K ATP channels in AgRP neurons are required for leptin regulation of body weight and blood glucose a , Schematic diagram of AAV pU6-sgRNA Kcnj11 ::pEF1α-FLEX-mCherry (AAV-sgK ATP ) unilaterally injected into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP mice and mCherry::GFP co-immunostaining. b , Representative traces of whole-cell current-clamp recordings in AgRP neurons from non-virus-transduced (gray, AAV − ) and virus-transduced sides (green, AAV + ) in 24-hour fasted Agrp-IRES-Cre::LSL-Cas9-GFP mice. K ATP channel opener diazoxide was applied as indicated ( n =12 neurons per group from 4 mice). c , d , mCherry::Fos co-immunostaining and representative traces of brain slice whole-cell current-clamp recordings in AgRP neurons of Agrp-IRES-Cre::LSL-Cas9-GFP mice with unilaterally-injected virus into the ARC ( c ), and quantification of frequency and membrane potential ( d ) ( n =10 neurons per group from 3 mice). e–n , Schematic diagram of AAV-sgK ATP bilaterally injected into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP mice and mCherry::GFP co-immunostaining ( e ), representative littermates ( f ), serum leptin levels ( g ), body weight ( h ), fat mass ( i ), daily food intake ( j ), ad libitum fed state blood glucose levels ( k ), serum insulin levels ( l ), Glucose-tolerance test ( m ), and Insulin-tolerance test ( n ) at 8 weeks post-viral injection ( n =9 mice per group). o , p , Body weight and daily food intake changes during 3-day consecutive treatment with saline/leptin ( n =9 mice per group). Data are mean ± s.e.m. and representative of three independent experiments; * P

    Journal: Nature

    Article Title: Genetic identification of leptin neural circuits in energy and glucose homeostases

    doi: 10.1038/s41586-018-0049-7

    Figure Lengend Snippet: K ATP channels in AgRP neurons are required for leptin regulation of body weight and blood glucose a , Schematic diagram of AAV pU6-sgRNA Kcnj11 ::pEF1α-FLEX-mCherry (AAV-sgK ATP ) unilaterally injected into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP mice and mCherry::GFP co-immunostaining. b , Representative traces of whole-cell current-clamp recordings in AgRP neurons from non-virus-transduced (gray, AAV − ) and virus-transduced sides (green, AAV + ) in 24-hour fasted Agrp-IRES-Cre::LSL-Cas9-GFP mice. K ATP channel opener diazoxide was applied as indicated ( n =12 neurons per group from 4 mice). c , d , mCherry::Fos co-immunostaining and representative traces of brain slice whole-cell current-clamp recordings in AgRP neurons of Agrp-IRES-Cre::LSL-Cas9-GFP mice with unilaterally-injected virus into the ARC ( c ), and quantification of frequency and membrane potential ( d ) ( n =10 neurons per group from 3 mice). e–n , Schematic diagram of AAV-sgK ATP bilaterally injected into the ARC of Agrp-IRES-Cre::LSL-Cas9-GFP mice and mCherry::GFP co-immunostaining ( e ), representative littermates ( f ), serum leptin levels ( g ), body weight ( h ), fat mass ( i ), daily food intake ( j ), ad libitum fed state blood glucose levels ( k ), serum insulin levels ( l ), Glucose-tolerance test ( m ), and Insulin-tolerance test ( n ) at 8 weeks post-viral injection ( n =9 mice per group). o , p , Body weight and daily food intake changes during 3-day consecutive treatment with saline/leptin ( n =9 mice per group). Data are mean ± s.e.m. and representative of three independent experiments; * P

    Article Snippet: Primary antibodies used in the current study and their dilutions are: rabbit anti-DsRed (Clontech, 1:2000, Cat#632496), chicken anti-mCherry (EnCor Biotechnology, 1:2000, # CPCA-mCherry), chicken anti-GFP (Aves Labs, 1:2000, GFP-1010), rabbit anti-hrGFP (Agilent Technologies, 1:1000, Cat# 240142), goat anti-Fos (Santa Cruz Biotechnology, 1:150, Cat# sc-52-g), rabbit anti-pSTAT3 (Cell Signaling Technology, 1:1000, Cat# 9145S), rabbit phosphor-S6 ribosomal protein (S235/236) (Cell Signaling Technology, 1:1000, Cat# 4858).

    Techniques: Injection, Mouse Assay, Immunostaining, Slice Preparation

    Characterization of feeding behaviors in STZ-treated diabetic mice; Additional analyses of ectopic activation of AgRP neurons and its pathologic contributions to STZ-induced hyperphagia and hyperglycemia a , Quantitative PCR results showing that Agrp and Npy mRNA levels are significantly upregulated in the mediobasal hypothalamus of STZ-treated animals, which is consistent with increased AgRP neuronal activity following STZ-injection ( n =5 mice per group). b–f , Food intake during light cycle (7 a.m. to 7 p.m.) ( b ) and dark cycle (7 p.m. to 7 a.m.) ( c ), representative 1 -hour heat map (10 to 11 a.m.) showing percent occupancy time in food zone (upper right corner) and nesting zone (lower right corner) ( d ), feeding duration ( e ), and 1-hour food intake ( f ) in saline- or STZ-treated C57BL/6 mice ( n =8 mice per group). g , h , Representative sections and quantification of hrGFP immunostaining in the ARC of saline- or STZ-treated Npy-hrGFP transgenic mice, suggesting that STZ-treatment does not induce obvious cell loss of AgRP neurons ( n =3 mice per group). i , Representative sections and quantification of hrGFP::pS6 co-immunostaining in the ARC of saline- or STZ-treated Npy-hrGFP transgenic mice ( n =4 mice per group). j , Schematic diagram of chemogenetic inhibition of AgRP neurons in virus-transduced Agrp-IRES-Cre mice. k , 4-hour food intake measurement during dark cycle (8 p.m.–12 a.m.) following the administration of saline or CNO ( n =6 mice per group). l , m , 4-hour food intake assay (10 a.m.–2 p.m.) ( l ) and blood glucose measurement ( m , without food in the cage) in female Agrp-IRES-Cre littermates after AAV pSyn-FLEX-hM4D i -mCherry virus injection into the ARC and upon saline/CNO treatment ( n =8 mice per group). n–q , Schematic diagram of experiments to assess CNO’s effects with Cre-dependent AAV-FLEX-mCherry virus injected to into the ARC of Agrp-IRES-Cre mice ( n ). Representative brain sections ( o ) and quantification ( p , n =3 mice per group) of mCherry::Fos co-immunostaining, and food intake assay ( q , 10 a.m. to 2 p.m., n =8 mice per group) in STZ-treated mice following the i.p. injection of saline/CNO, demonstrating that CNO administration without the expression of hM 4 Di in AgRP neurons induces null effects and the changes observed in are caused by the chemogenetic inhibition of AgRP neurons. Data are mean ± s.e.m. and representative of three independent experiments; * P

    Journal: Nature

    Article Title: Genetic identification of leptin neural circuits in energy and glucose homeostases

    doi: 10.1038/s41586-018-0049-7

    Figure Lengend Snippet: Characterization of feeding behaviors in STZ-treated diabetic mice; Additional analyses of ectopic activation of AgRP neurons and its pathologic contributions to STZ-induced hyperphagia and hyperglycemia a , Quantitative PCR results showing that Agrp and Npy mRNA levels are significantly upregulated in the mediobasal hypothalamus of STZ-treated animals, which is consistent with increased AgRP neuronal activity following STZ-injection ( n =5 mice per group). b–f , Food intake during light cycle (7 a.m. to 7 p.m.) ( b ) and dark cycle (7 p.m. to 7 a.m.) ( c ), representative 1 -hour heat map (10 to 11 a.m.) showing percent occupancy time in food zone (upper right corner) and nesting zone (lower right corner) ( d ), feeding duration ( e ), and 1-hour food intake ( f ) in saline- or STZ-treated C57BL/6 mice ( n =8 mice per group). g , h , Representative sections and quantification of hrGFP immunostaining in the ARC of saline- or STZ-treated Npy-hrGFP transgenic mice, suggesting that STZ-treatment does not induce obvious cell loss of AgRP neurons ( n =3 mice per group). i , Representative sections and quantification of hrGFP::pS6 co-immunostaining in the ARC of saline- or STZ-treated Npy-hrGFP transgenic mice ( n =4 mice per group). j , Schematic diagram of chemogenetic inhibition of AgRP neurons in virus-transduced Agrp-IRES-Cre mice. k , 4-hour food intake measurement during dark cycle (8 p.m.–12 a.m.) following the administration of saline or CNO ( n =6 mice per group). l , m , 4-hour food intake assay (10 a.m.–2 p.m.) ( l ) and blood glucose measurement ( m , without food in the cage) in female Agrp-IRES-Cre littermates after AAV pSyn-FLEX-hM4D i -mCherry virus injection into the ARC and upon saline/CNO treatment ( n =8 mice per group). n–q , Schematic diagram of experiments to assess CNO’s effects with Cre-dependent AAV-FLEX-mCherry virus injected to into the ARC of Agrp-IRES-Cre mice ( n ). Representative brain sections ( o ) and quantification ( p , n =3 mice per group) of mCherry::Fos co-immunostaining, and food intake assay ( q , 10 a.m. to 2 p.m., n =8 mice per group) in STZ-treated mice following the i.p. injection of saline/CNO, demonstrating that CNO administration without the expression of hM 4 Di in AgRP neurons induces null effects and the changes observed in are caused by the chemogenetic inhibition of AgRP neurons. Data are mean ± s.e.m. and representative of three independent experiments; * P

    Article Snippet: Primary antibodies used in the current study and their dilutions are: rabbit anti-DsRed (Clontech, 1:2000, Cat#632496), chicken anti-mCherry (EnCor Biotechnology, 1:2000, # CPCA-mCherry), chicken anti-GFP (Aves Labs, 1:2000, GFP-1010), rabbit anti-hrGFP (Agilent Technologies, 1:1000, Cat# 240142), goat anti-Fos (Santa Cruz Biotechnology, 1:150, Cat# sc-52-g), rabbit anti-pSTAT3 (Cell Signaling Technology, 1:1000, Cat# 9145S), rabbit phosphor-S6 ribosomal protein (S235/236) (Cell Signaling Technology, 1:1000, Cat# 4858).

    Techniques: Mouse Assay, Activation Assay, Real-time Polymerase Chain Reaction, Activity Assay, Injection, Immunostaining, Transgenic Assay, Inhibition, Expressing

    CRISPR-mediated disruption of leptin receptors in the hypothalamic POMC neurons does not alter energy or blood glucose balances a , b , Representative sections and quantification of hrGFP::Fos co-immunostaining in the ARC of saline- or STZ-treated Pomc-hrGFP transgenic mice ( n =3 mice per group). Reduced Fos expression in POMC neurons was observed following STZ treatment. c , Quantitative PCR results showing that Pomc mRNA levels are significantly reduced in the mediobasal hypothalamus of STZ-treated animals, which is consistent with inhibited POMC neuronal activities following STZ-injection ( n =5 mice per group). d , Schematic diagrams, representative sections, and quantification of tdTomato::hrGFP co-immunostaining in the ARC of Pomc-Cre::LSL-tdTomato::Npy-hrGFP mice (I, upper panels) and of mCherry::hrGFP co-immunostaining in the ARC of Pomc-Cre::Npy-hrGFP mice with Cre-dependent AAV pEF1α-FLEX-mCherry injected into the ARC (II, lower panels). Considerable co-expression was observed in Pomc-Cre::LSL-tdTomato::Npy-hrGFP mice but not in virus-transduced Pomc-Cre::Npy-hrGFP mice, suggesting that Pomc-Cre ectopically express Cre activity in AgRP neurons during early developmental stage, consistent with prior findings. These data also demonstrate that Cre-dependent AAV injected into the ARC of Pomc-cre transgenic mice provides an efficient approach to specifically express gene-of-interest in POMC ARC neurons, without perturbing intermingled AgRP neurons ( n =3 mice per group). e , Schematic diagrams of two approaches to achieve CRISPR-mediated deletion of Lepr in POMC ARC neurons. (I) AAV-sgLepR virus was bilaterally injected into the ARC of Pomc-Cre::LSL-Cas9-GFP mice. (II) A viral mix of AAV-sgLepR and Cre-dependent AAV pMeCP2-FLEX-spCas9 (AAV-FLEX-spCas9, see online methods for the details) was bilaterally injected into the ARC of Pomc-Cre mice. f–j , Body weight ( f ), accumulated weekly food intake ( g ), ad libitum fed state blood glucose levels ( h ), Glucose-tolerance test ( i ), and Insulin-tolerance test ( j ) of the virus-transduced animals ( n =6 mice per group). Of note, mildly-increased body weight and food intake, as well as slightly-impaired glucose tolerance and insulin sensitivity, were observed only in single virus-transduced Pomc-Cre::LSL-Cas9-GFP mice, but not in dual virus-transduced Pomc-Cre mice. Since Cas9 protein is expected to express in some AgRP neurons in Pomc-Cre::LSL-Cas9-GFP animals, the difference observed between the two approaches was likely explained by the ectopic Cre-activity of Pomc-Cre in AgRP neurons. The phenotypes observed in Pomc-Cre::LSL-Cas9-GFP mice mimic those following genetic ablation of Lepr with conventional Cre-loxP system. Inactivation of LepR in POMC neurons of adult mice does not appear to affect energy balance or glucose homeostasis under the conditions assayed. Data are mean ± s.e.m. and representative of three independent experiments; *P

    Journal: Nature

    Article Title: Genetic identification of leptin neural circuits in energy and glucose homeostases

    doi: 10.1038/s41586-018-0049-7

    Figure Lengend Snippet: CRISPR-mediated disruption of leptin receptors in the hypothalamic POMC neurons does not alter energy or blood glucose balances a , b , Representative sections and quantification of hrGFP::Fos co-immunostaining in the ARC of saline- or STZ-treated Pomc-hrGFP transgenic mice ( n =3 mice per group). Reduced Fos expression in POMC neurons was observed following STZ treatment. c , Quantitative PCR results showing that Pomc mRNA levels are significantly reduced in the mediobasal hypothalamus of STZ-treated animals, which is consistent with inhibited POMC neuronal activities following STZ-injection ( n =5 mice per group). d , Schematic diagrams, representative sections, and quantification of tdTomato::hrGFP co-immunostaining in the ARC of Pomc-Cre::LSL-tdTomato::Npy-hrGFP mice (I, upper panels) and of mCherry::hrGFP co-immunostaining in the ARC of Pomc-Cre::Npy-hrGFP mice with Cre-dependent AAV pEF1α-FLEX-mCherry injected into the ARC (II, lower panels). Considerable co-expression was observed in Pomc-Cre::LSL-tdTomato::Npy-hrGFP mice but not in virus-transduced Pomc-Cre::Npy-hrGFP mice, suggesting that Pomc-Cre ectopically express Cre activity in AgRP neurons during early developmental stage, consistent with prior findings. These data also demonstrate that Cre-dependent AAV injected into the ARC of Pomc-cre transgenic mice provides an efficient approach to specifically express gene-of-interest in POMC ARC neurons, without perturbing intermingled AgRP neurons ( n =3 mice per group). e , Schematic diagrams of two approaches to achieve CRISPR-mediated deletion of Lepr in POMC ARC neurons. (I) AAV-sgLepR virus was bilaterally injected into the ARC of Pomc-Cre::LSL-Cas9-GFP mice. (II) A viral mix of AAV-sgLepR and Cre-dependent AAV pMeCP2-FLEX-spCas9 (AAV-FLEX-spCas9, see online methods for the details) was bilaterally injected into the ARC of Pomc-Cre mice. f–j , Body weight ( f ), accumulated weekly food intake ( g ), ad libitum fed state blood glucose levels ( h ), Glucose-tolerance test ( i ), and Insulin-tolerance test ( j ) of the virus-transduced animals ( n =6 mice per group). Of note, mildly-increased body weight and food intake, as well as slightly-impaired glucose tolerance and insulin sensitivity, were observed only in single virus-transduced Pomc-Cre::LSL-Cas9-GFP mice, but not in dual virus-transduced Pomc-Cre mice. Since Cas9 protein is expected to express in some AgRP neurons in Pomc-Cre::LSL-Cas9-GFP animals, the difference observed between the two approaches was likely explained by the ectopic Cre-activity of Pomc-Cre in AgRP neurons. The phenotypes observed in Pomc-Cre::LSL-Cas9-GFP mice mimic those following genetic ablation of Lepr with conventional Cre-loxP system. Inactivation of LepR in POMC neurons of adult mice does not appear to affect energy balance or glucose homeostasis under the conditions assayed. Data are mean ± s.e.m. and representative of three independent experiments; *P

    Article Snippet: Primary antibodies used in the current study and their dilutions are: rabbit anti-DsRed (Clontech, 1:2000, Cat#632496), chicken anti-mCherry (EnCor Biotechnology, 1:2000, # CPCA-mCherry), chicken anti-GFP (Aves Labs, 1:2000, GFP-1010), rabbit anti-hrGFP (Agilent Technologies, 1:1000, Cat# 240142), goat anti-Fos (Santa Cruz Biotechnology, 1:150, Cat# sc-52-g), rabbit anti-pSTAT3 (Cell Signaling Technology, 1:1000, Cat# 9145S), rabbit phosphor-S6 ribosomal protein (S235/236) (Cell Signaling Technology, 1:1000, Cat# 4858).

    Techniques: CRISPR, Immunostaining, Transgenic Assay, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction, Injection, Activity Assay

    Dynamic changes of neuronal activity and synaptic neurotransmission following CRISPR-mediated deletion of GABA A receptors in AgRP neurons a , b , Body weight measurement ( a ) and daily food intake ( b ) of Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice following bilateral injection of AAV-sgGABA A -R into the ARC ( n =7 mice per group). c–f , Representative sections and quantification of mCherry::Fos co-immunostaining ( c , n =3 mice per group), representative traces and quantification of spontaneous excitatory postsynaptic currents (sEPSCs) ( d , n =15 neurons from 3 mice, e , n =8 neurons from 2 mice) in AgRP neurons of Agrp-IRES-Cre::LSL-Cas9-GFP mice after 1-week or 4-week unilateral injection of AAV-sgGABA A -R into the ARC. Reduced frequency and amplitude of sEPSCs were observed in AgRP neurons in a later stage following GABA A -R deletion, together with concurrent elimination of the observed disinhibition of these neurons ( c ), body weight gain ( a ), and hyperphagia ( b ), suggesting that excitatory and inhibitory afferents dynamically cooperate to modulate AgRP neuronal activities. In addition, the compensatory reduction in sEPSCs in virus-transduced neurons but not in the neurons from the contralateral side also indicate a cell-autonomous mechanism. Data are mean ± s.e.m. and representative of three independent experiments; **P

    Journal: Nature

    Article Title: Genetic identification of leptin neural circuits in energy and glucose homeostases

    doi: 10.1038/s41586-018-0049-7

    Figure Lengend Snippet: Dynamic changes of neuronal activity and synaptic neurotransmission following CRISPR-mediated deletion of GABA A receptors in AgRP neurons a , b , Body weight measurement ( a ) and daily food intake ( b ) of Agrp-IRES-Cre (Cas9 − ) and Agrp-IRES-Cre::LSL-Cas9-GFP (Cas9 + ) mice following bilateral injection of AAV-sgGABA A -R into the ARC ( n =7 mice per group). c–f , Representative sections and quantification of mCherry::Fos co-immunostaining ( c , n =3 mice per group), representative traces and quantification of spontaneous excitatory postsynaptic currents (sEPSCs) ( d , n =15 neurons from 3 mice, e , n =8 neurons from 2 mice) in AgRP neurons of Agrp-IRES-Cre::LSL-Cas9-GFP mice after 1-week or 4-week unilateral injection of AAV-sgGABA A -R into the ARC. Reduced frequency and amplitude of sEPSCs were observed in AgRP neurons in a later stage following GABA A -R deletion, together with concurrent elimination of the observed disinhibition of these neurons ( c ), body weight gain ( a ), and hyperphagia ( b ), suggesting that excitatory and inhibitory afferents dynamically cooperate to modulate AgRP neuronal activities. In addition, the compensatory reduction in sEPSCs in virus-transduced neurons but not in the neurons from the contralateral side also indicate a cell-autonomous mechanism. Data are mean ± s.e.m. and representative of three independent experiments; **P

    Article Snippet: Primary antibodies used in the current study and their dilutions are: rabbit anti-DsRed (Clontech, 1:2000, Cat#632496), chicken anti-mCherry (EnCor Biotechnology, 1:2000, # CPCA-mCherry), chicken anti-GFP (Aves Labs, 1:2000, GFP-1010), rabbit anti-hrGFP (Agilent Technologies, 1:1000, Cat# 240142), goat anti-Fos (Santa Cruz Biotechnology, 1:150, Cat# sc-52-g), rabbit anti-pSTAT3 (Cell Signaling Technology, 1:1000, Cat# 9145S), rabbit phosphor-S6 ribosomal protein (S235/236) (Cell Signaling Technology, 1:1000, Cat# 4858).

    Techniques: Activity Assay, CRISPR, Mouse Assay, Injection, Immunostaining

    Overactivation of AgRP neurons induces diabetic hyperphagia and hyperglycemia a , b , Fos immunostaining in the mediobasal hypothalamus of STZ-treated C57BL/6 mice 3 hours after the administration of saline/leptin ( n =3 mice per group). ARC, arcuate nucleus; DMH, dorsomedial hypothalamus; LH, lateral hypothalamus; 3V, 3 rd ventricle. c , d , Fos::hrGFP co-immunostaining in the ARC of Npy-hrGFP transgenic mice following saline/STZ treatment ( n =4 mice per group). e , Representative traces of brain slice whole-cell current clamp recordings in AgRP neurons of Npy-hrGFP mice treated with saline/STZ ( n =10 neurons per group from 3 mice). f , Diagram of AAV pSyn-FLEX-hM4D i -mCherry injected into the ARC of STZ-treated Agrp-IRES-Cre mice and mCherry immunostaining. g , h , mCherry::Fos co-immunostaining in STZ-treated Agrp-IRES-Cre mice following the administration of saline/CNO ( n =4 mice per group). i , Food intake (10 a.m.-2 p.m.) in ad libitum fed, virus-transduced Agrp-IRES-Cre mice following the administration of saline/CNO ( n =8 mice per group). j , k , Experimental design and blood glucose before and 1 hour after Saline/CNO treatment in STZ-treated, virus-transduced Agrp-IRES-Cre mice ( n =8 mice per group). Data are mean ± s.e.m. and representative of three independent experiments; ** P

    Journal: Nature

    Article Title: Genetic identification of leptin neural circuits in energy and glucose homeostases

    doi: 10.1038/s41586-018-0049-7

    Figure Lengend Snippet: Overactivation of AgRP neurons induces diabetic hyperphagia and hyperglycemia a , b , Fos immunostaining in the mediobasal hypothalamus of STZ-treated C57BL/6 mice 3 hours after the administration of saline/leptin ( n =3 mice per group). ARC, arcuate nucleus; DMH, dorsomedial hypothalamus; LH, lateral hypothalamus; 3V, 3 rd ventricle. c , d , Fos::hrGFP co-immunostaining in the ARC of Npy-hrGFP transgenic mice following saline/STZ treatment ( n =4 mice per group). e , Representative traces of brain slice whole-cell current clamp recordings in AgRP neurons of Npy-hrGFP mice treated with saline/STZ ( n =10 neurons per group from 3 mice). f , Diagram of AAV pSyn-FLEX-hM4D i -mCherry injected into the ARC of STZ-treated Agrp-IRES-Cre mice and mCherry immunostaining. g , h , mCherry::Fos co-immunostaining in STZ-treated Agrp-IRES-Cre mice following the administration of saline/CNO ( n =4 mice per group). i , Food intake (10 a.m.-2 p.m.) in ad libitum fed, virus-transduced Agrp-IRES-Cre mice following the administration of saline/CNO ( n =8 mice per group). j , k , Experimental design and blood glucose before and 1 hour after Saline/CNO treatment in STZ-treated, virus-transduced Agrp-IRES-Cre mice ( n =8 mice per group). Data are mean ± s.e.m. and representative of three independent experiments; ** P

    Article Snippet: Primary antibodies used in the current study and their dilutions are: rabbit anti-DsRed (Clontech, 1:2000, Cat#632496), chicken anti-mCherry (EnCor Biotechnology, 1:2000, # CPCA-mCherry), chicken anti-GFP (Aves Labs, 1:2000, GFP-1010), rabbit anti-hrGFP (Agilent Technologies, 1:1000, Cat# 240142), goat anti-Fos (Santa Cruz Biotechnology, 1:150, Cat# sc-52-g), rabbit anti-pSTAT3 (Cell Signaling Technology, 1:1000, Cat# 9145S), rabbit phosphor-S6 ribosomal protein (S235/236) (Cell Signaling Technology, 1:1000, Cat# 4858).

    Techniques: Immunostaining, Mouse Assay, Transgenic Assay, Slice Preparation, Injection

    Id + diffuse large B cell lymphomas are infiltrated with Id-specific T cells. (A) Lymphomatous spleen of an Id-Th2→Id + mouse (top) compared with spleen from an Ova-Th2→Id + mouse (bottom). (B) Photo of enlarged mesenteric LN in an Id-Th2→Id + mouse with lymphoma (left). The circled asterisk indicates the normal size of a mesenteric LN in an Ova-Th2→Id + mouse. (right) Liver from Id-Th2→Id + mouse. Arrows indicate tumor nodules. (C) Low and high power magnification (hematoxylin and eosin) of lymphomatous spleen. (D) Lymphoma of the liver with infiltrating CD3 + T cells (brown). (E) Spleen lymphoma were B220 + , IRF-4/MUM1 + , and CD138 + (in about half of the mice) and were IgM + IgD + . In the rightmost immunofluorescence picture, DAPI stains the nuclei. (F) Low power magnification of splenic lymphoma, Id + λ2 L chain (red, 2B6 mAb) versus κ (green). (inset) Large Id + lymphoma cell compared with smaller κ + B cell. (G) Id-specific T cells (red, TCR clonotype-specific mAb GB113) and lymphoma cells (green, 2B6 mAb) were often in close contact (yellow). See inset for higher magnification of a synapse.

    Journal: The Journal of Experimental Medicine

    Article Title: Lymphomas can develop from B cells chronically helped by idiotype-specific T cells

    doi: 10.1084/jem.20061220

    Figure Lengend Snippet: Id + diffuse large B cell lymphomas are infiltrated with Id-specific T cells. (A) Lymphomatous spleen of an Id-Th2→Id + mouse (top) compared with spleen from an Ova-Th2→Id + mouse (bottom). (B) Photo of enlarged mesenteric LN in an Id-Th2→Id + mouse with lymphoma (left). The circled asterisk indicates the normal size of a mesenteric LN in an Ova-Th2→Id + mouse. (right) Liver from Id-Th2→Id + mouse. Arrows indicate tumor nodules. (C) Low and high power magnification (hematoxylin and eosin) of lymphomatous spleen. (D) Lymphoma of the liver with infiltrating CD3 + T cells (brown). (E) Spleen lymphoma were B220 + , IRF-4/MUM1 + , and CD138 + (in about half of the mice) and were IgM + IgD + . In the rightmost immunofluorescence picture, DAPI stains the nuclei. (F) Low power magnification of splenic lymphoma, Id + λ2 L chain (red, 2B6 mAb) versus κ (green). (inset) Large Id + lymphoma cell compared with smaller κ + B cell. (G) Id-specific T cells (red, TCR clonotype-specific mAb GB113) and lymphoma cells (green, 2B6 mAb) were often in close contact (yellow). See inset for higher magnification of a synapse.

    Article Snippet: Primary rabbit anti-CD3 antibody (DakoCytomation), rat-anti B220 (clone RA3-6B2, BD Biosciences), goat anti-MUM1/IRF-4 (M-17; Santa Cruz Biotechnology, Inc.), rabbit anti–BCL-6 (N-3; Santa Cruz Biotechnology, Inc.), or rat anti-CD138 (clone 281-2; BD Biosciences) was applied in DakoCytomation diluent for 1 h. For B220 and CD138, rabbit anti–rat Ig antibody (DakoCytomation) was applied in DakoCytomation diluent for 1 h. Slides were washed in 50 mM Tris-Cl, pH 7.4, and detected with anti-rabbit Envision+ kit (CD3, B220, BCL-6, and CD138; DakoCytomation) or LSAB+ staining kit (MUM1/IRF-4; DakoCytomation) as per the manufacturer's instructions.

    Techniques: Mouse Assay, Immunofluorescence

    N terminus and first transmembrane segment of Na v 1.6 are sufficient to direct a reporter protein to the cell surface. Confocal images of HEK293 cells transfected with the reporter constructs and probed with anti-CD4 antibodies are shown. Green , anti-CD74; blue , DAPI. A , extracellular domain of CD74 lacking membrane-targeting N terminus. B , N terminus and first transmembrane segment of Na v 1.6 fused to the extracellular domain of CD74. C , ataxia3 mutation S21P not disrupting surface localization.

    Journal: The Journal of Biological Chemistry

    Article Title: Interaction of Voltage-gated Sodium Channel Nav1.6 (SCN8A) with Microtubule-associated Protein Map1b *

    doi: 10.1074/jbc.M111.336024

    Figure Lengend Snippet: N terminus and first transmembrane segment of Na v 1.6 are sufficient to direct a reporter protein to the cell surface. Confocal images of HEK293 cells transfected with the reporter constructs and probed with anti-CD4 antibodies are shown. Green , anti-CD74; blue , DAPI. A , extracellular domain of CD74 lacking membrane-targeting N terminus. B , N terminus and first transmembrane segment of Na v 1.6 fused to the extracellular domain of CD74. C , ataxia3 mutation S21P not disrupting surface localization.

    Article Snippet: Cells were blocked in 10% donkey serum in PBS (D9663; Sigma) and incubated at 4 °C overnight with a 1:750 dilution of anti-CD74 (CD74 (C-16) goat polyclonal IgG, sc-5438; Santa Cruz Biotechnology) in 20% donkey serum-PBS.

    Techniques: Transfection, Construct, Mutagenesis

    Interaction of Na v 1.6 with Map1b. A , HEK293 cells co-transfected with Na v 1.6-CD74 and myc-Map1b. Map1b light chain co-immunoprecipitates with Na v 1.6-CD74 fusion protein from co-transfected cells. B , voltage-gated sodium channels and Map1b light chain co-immunoprecipitated from brain membrane fractions from wild-type (+/+) but not Na v 1.6 null (−/−) mice. Lanes contain 50 μg of protein (brain) or 100 μg of protein (immunoprecipitate).

    Journal: The Journal of Biological Chemistry

    Article Title: Interaction of Voltage-gated Sodium Channel Nav1.6 (SCN8A) with Microtubule-associated Protein Map1b *

    doi: 10.1074/jbc.M111.336024

    Figure Lengend Snippet: Interaction of Na v 1.6 with Map1b. A , HEK293 cells co-transfected with Na v 1.6-CD74 and myc-Map1b. Map1b light chain co-immunoprecipitates with Na v 1.6-CD74 fusion protein from co-transfected cells. B , voltage-gated sodium channels and Map1b light chain co-immunoprecipitated from brain membrane fractions from wild-type (+/+) but not Na v 1.6 null (−/−) mice. Lanes contain 50 μg of protein (brain) or 100 μg of protein (immunoprecipitate).

    Article Snippet: Cells were blocked in 10% donkey serum in PBS (D9663; Sigma) and incubated at 4 °C overnight with a 1:750 dilution of anti-CD74 (CD74 (C-16) goat polyclonal IgG, sc-5438; Santa Cruz Biotechnology) in 20% donkey serum-PBS.

    Techniques: Transfection, Immunoprecipitation, Mouse Assay

    HCt/E-specific antibody production in mice immunized with BoNT/HCt/E DNA vaccine by intradermal electroporation. Mice were immunized 3 times (week 0, 2, 4) with 50 μg of DNA vaccine. Anti-HCt/E IgG levels are expressed as endpoint titers. Each symbol represents data from an individual mouse. The bar represents the mean titer for each group.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Intradermal immunization with botulinum neurotoxin serotype E DNA vaccine induces humoral and cellular immunity and protects against lethal toxin challenge

    doi: 10.1080/21645515.2018.1526554

    Figure Lengend Snippet: HCt/E-specific antibody production in mice immunized with BoNT/HCt/E DNA vaccine by intradermal electroporation. Mice were immunized 3 times (week 0, 2, 4) with 50 μg of DNA vaccine. Anti-HCt/E IgG levels are expressed as endpoint titers. Each symbol represents data from an individual mouse. The bar represents the mean titer for each group.

    Article Snippet: The HCt/E protein was detected using a mouse anti-HCt/E antibody and goat anti-mouse IgG conjugated to horseradish peroxidase (Santa Cruz Biotech, Santa Cruz, USA).

    Techniques: Mouse Assay, Electroporation

    Survival and antibody titers following immunization with HCt/E protein vaccine. Mice were immunized three times at 2-week intervals with 10 μg or 20 μg of purified HCt/E vaccine intramuscularly, followed by intraperitoneal challenge with 200 LD 50 of active BoNT/E. The number of survivors are shown. Anti-HCt/E IgG levels are expressed as endpoint titers. Sera were obtained one week after last immunization. Each symbol represents data from an individual mouse. The bar represents the mean titer for each group.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Intradermal immunization with botulinum neurotoxin serotype E DNA vaccine induces humoral and cellular immunity and protects against lethal toxin challenge

    doi: 10.1080/21645515.2018.1526554

    Figure Lengend Snippet: Survival and antibody titers following immunization with HCt/E protein vaccine. Mice were immunized three times at 2-week intervals with 10 μg or 20 μg of purified HCt/E vaccine intramuscularly, followed by intraperitoneal challenge with 200 LD 50 of active BoNT/E. The number of survivors are shown. Anti-HCt/E IgG levels are expressed as endpoint titers. Sera were obtained one week after last immunization. Each symbol represents data from an individual mouse. The bar represents the mean titer for each group.

    Article Snippet: The HCt/E protein was detected using a mouse anti-HCt/E antibody and goat anti-mouse IgG conjugated to horseradish peroxidase (Santa Cruz Biotech, Santa Cruz, USA).

    Techniques: Mouse Assay, Purification

    Survival and antibody titers following immunization with BoNT/HCt/E DNA vaccine. Mice were immunized two or three times at 2-week intervals with various doses of DNA vaccine intradermally via electroporation, followed by intraperitoneal challenge with 50 LD 50 of active BoNT/E. (A) Survival curves show the percentage of mice alive after challenge. Data shown are from one of two experiments. (B) Anti-HCt/E IgG levels are expressed as endpoint titers. Sera were obtained one week before challenged. Each symbol represents data from an individual mouse. The bar represents the mean titer for each group. Control group mice were immunized with vector only. Significant results are marked asterisks: ** p

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Intradermal immunization with botulinum neurotoxin serotype E DNA vaccine induces humoral and cellular immunity and protects against lethal toxin challenge

    doi: 10.1080/21645515.2018.1526554

    Figure Lengend Snippet: Survival and antibody titers following immunization with BoNT/HCt/E DNA vaccine. Mice were immunized two or three times at 2-week intervals with various doses of DNA vaccine intradermally via electroporation, followed by intraperitoneal challenge with 50 LD 50 of active BoNT/E. (A) Survival curves show the percentage of mice alive after challenge. Data shown are from one of two experiments. (B) Anti-HCt/E IgG levels are expressed as endpoint titers. Sera were obtained one week before challenged. Each symbol represents data from an individual mouse. The bar represents the mean titer for each group. Control group mice were immunized with vector only. Significant results are marked asterisks: ** p

    Article Snippet: The HCt/E protein was detected using a mouse anti-HCt/E antibody and goat anti-mouse IgG conjugated to horseradish peroxidase (Santa Cruz Biotech, Santa Cruz, USA).

    Techniques: Mouse Assay, Electroporation, Plasmid Preparation