anti human kv1 3  (Jackson Immuno)

 
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    Name:
    Rhodamine TRITC AffiniPure Donkey Anti Chicken IgY IgG
    Description:
    Whole IgG antibodies are isolated as intact molecules from antisera by immunoaffinity chromatography They have an Fc portion and two antigen binding Fab portions joined together by disulfide bonds and therefore they are divalent The average molecular weight is reported to be about 160 kDa The whole IgG form of antibodies is suitable for the majority of immunodetection procedures and is the most cost effective Based on immunoelectrophoresis and or ELISA the antibody reacts with whole molecule chicken IgY It also reacts with the light chains of other chicken immunoglobulins No antibody was detected against non immunoglobulin serum proteins The antibody has been tested by ELISA and or solid phase adsorbed to ensure minimal cross reaction with bovine goat guinea pig syrian hamster horse human mouse rabbit rat and sheep serum proteins but it may cross react with immunoglobulins from other species
    Catalog Number:
    703-025-155
    Price:
    97
    Purity:
    The antibody was purified from antisera by immunoaffinity chromatography using antigens coupled to agarose beads.
    Conjugate:
    Rhodamine TRITC
    Size:
    mg
    Category:
    Secondary Antibody
    Source:
    Donkey
    Quantity:
    0 5
    Buy from Supplier


    Structured Review

    Jackson Immuno anti human kv1 3
    Rhodamine TRITC AffiniPure Donkey Anti Chicken IgY IgG
    Whole IgG antibodies are isolated as intact molecules from antisera by immunoaffinity chromatography They have an Fc portion and two antigen binding Fab portions joined together by disulfide bonds and therefore they are divalent The average molecular weight is reported to be about 160 kDa The whole IgG form of antibodies is suitable for the majority of immunodetection procedures and is the most cost effective Based on immunoelectrophoresis and or ELISA the antibody reacts with whole molecule chicken IgY It also reacts with the light chains of other chicken immunoglobulins No antibody was detected against non immunoglobulin serum proteins The antibody has been tested by ELISA and or solid phase adsorbed to ensure minimal cross reaction with bovine goat guinea pig syrian hamster horse human mouse rabbit rat and sheep serum proteins but it may cross react with immunoglobulins from other species
    https://www.bioz.com/result/anti human kv1 3/product/Jackson Immuno
    Average 85 stars, based on 351 article reviews
    Price from $9.99 to $1999.99
    anti human kv1 3 - by Bioz Stars, 2020-09
    85/100 stars

    Images

    1) Product Images from "The Ca2+-Activated K+ Channel KCa3.1 as a Potential New Target for the Prevention of Allograft Vasculopathy"

    Article Title: The Ca2+-Activated K+ Channel KCa3.1 as a Potential New Target for the Prevention of Allograft Vasculopathy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0081006

    Kv1.3 and KCa3.1 expression in human and rat vasculopathy. (A, B) KCa3.1 and Kv1.3 staining in serial sections from a coronary artery with severe atherosclerotic changes. The vessel was harvested from the former heart of a patient receiving a heart transplant because of ischemic cardiomyopathy. (C, D) KCa3.1 and Kv1.3 staining in a mammary artery bypass graft. (E, F) KCa3.1 staining in orthotopic rat aorta transplants harvested 80 or 120 days after transplantation. G, Close-up of the boxed area in F. (H, I) Kv1.3 staining in serial sections of the grafts shown in E and F. (J) Close-up of the boxed area in F. Serial sections are 5 µm apart.
    Figure Legend Snippet: Kv1.3 and KCa3.1 expression in human and rat vasculopathy. (A, B) KCa3.1 and Kv1.3 staining in serial sections from a coronary artery with severe atherosclerotic changes. The vessel was harvested from the former heart of a patient receiving a heart transplant because of ischemic cardiomyopathy. (C, D) KCa3.1 and Kv1.3 staining in a mammary artery bypass graft. (E, F) KCa3.1 staining in orthotopic rat aorta transplants harvested 80 or 120 days after transplantation. G, Close-up of the boxed area in F. (H, I) Kv1.3 staining in serial sections of the grafts shown in E and F. (J) Close-up of the boxed area in F. Serial sections are 5 µm apart.

    Techniques Used: Expressing, Staining, Transplantation Assay

    2) Product Images from "BMAL1 Shuttling Controls Transactivation and Degradation of the CLOCK/BMAL1 Heterodimer"

    Article Title: BMAL1 Shuttling Controls Transactivation and Degradation of the CLOCK/BMAL1 Heterodimer

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.00337-06

    BMAL1-dependent proteolysis of CLOCK is closely related to its transcriptional activity. (A) Intact BMAL1 but not functionally defective BMAL1 induces drastic decreases in CLOCK levels. NIH 3T3 cells were transfected with Myc-Clock alone or together with wild-type Bmal1 or Bmal1 mutants and then analyzed by immunoblotting with the indicated antibodies. (B) Confirmation of protein-protein interactions between CLOCK and BMAL1 mutants. Cells were cotransfected with wild-type CLOCK and Myc-tagged Bmal1 constructs with various defects and then immunoprecipitated with anti-Myc antibodies. The immunoprecipitates were analyzed by immunoblotting with anti-CLOCK or anti-Myc. (C) BMAL1-dependent CLOCK degradation is prevented when CLOCK has functional defects or is coexpressed with CRY1. Cells were transfected with Myc-tagged Clock constructs encoding wild-type or transcriptionally inactive mutants [ΔN (52-855) lacking the DNA-binding domain and ΔC (1-480) lacking TAD] alone or together with Bmal1 and/or Cry1 and then analyzed by immunoblotting with the indicated antibodies. (D) The CLOCK mutants lacking DNA-binding domain or TAD colocalize with BMAL1 in the nucleus. Localization of the Myc-tagged CLOCK mutants and BMAL1 was detected with anti-Myc and anti-BMAL1 antibodies and visualized with TRITC and FITC, respectively. ( E) Relative Per1 promoter-dependent luciferase reporter activity induced by combined transfection with the various forms of Clock and Bmal1. The rightmost bar is the result from cells transfected with wild-type Clock and Bmal1 and exposed to 50 μM MG132 for 5 h. Relative luciferase reporter activity is expressed as fold of the control. Each value is the mean ± the SEM of three replicates from a single assay. Similar results were obtained from replicated experiments.
    Figure Legend Snippet: BMAL1-dependent proteolysis of CLOCK is closely related to its transcriptional activity. (A) Intact BMAL1 but not functionally defective BMAL1 induces drastic decreases in CLOCK levels. NIH 3T3 cells were transfected with Myc-Clock alone or together with wild-type Bmal1 or Bmal1 mutants and then analyzed by immunoblotting with the indicated antibodies. (B) Confirmation of protein-protein interactions between CLOCK and BMAL1 mutants. Cells were cotransfected with wild-type CLOCK and Myc-tagged Bmal1 constructs with various defects and then immunoprecipitated with anti-Myc antibodies. The immunoprecipitates were analyzed by immunoblotting with anti-CLOCK or anti-Myc. (C) BMAL1-dependent CLOCK degradation is prevented when CLOCK has functional defects or is coexpressed with CRY1. Cells were transfected with Myc-tagged Clock constructs encoding wild-type or transcriptionally inactive mutants [ΔN (52-855) lacking the DNA-binding domain and ΔC (1-480) lacking TAD] alone or together with Bmal1 and/or Cry1 and then analyzed by immunoblotting with the indicated antibodies. (D) The CLOCK mutants lacking DNA-binding domain or TAD colocalize with BMAL1 in the nucleus. Localization of the Myc-tagged CLOCK mutants and BMAL1 was detected with anti-Myc and anti-BMAL1 antibodies and visualized with TRITC and FITC, respectively. ( E) Relative Per1 promoter-dependent luciferase reporter activity induced by combined transfection with the various forms of Clock and Bmal1. The rightmost bar is the result from cells transfected with wild-type Clock and Bmal1 and exposed to 50 μM MG132 for 5 h. Relative luciferase reporter activity is expressed as fold of the control. Each value is the mean ± the SEM of three replicates from a single assay. Similar results were obtained from replicated experiments.

    Techniques Used: Activity Assay, Transfection, Construct, Immunoprecipitation, Functional Assay, Binding Assay, Luciferase

    3) Product Images from "Production of Anti-carbohydrate Antibodies by Phage Display Technologies"

    Article Title: Production of Anti-carbohydrate Antibodies by Phage Display Technologies

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.107284

    Colocalization of scFv-Fc or Fc proteins with ER. Left panel , COS-7 cells transfected with scFv-Fc genes encoding 3F1, 1F12, 5A3, 1A4, 1G4, or Fc and cultured for 16 h were fixed. Fixed cells were stained with anti-PDI antibody and FITC-conjugated AffiniPure donkey anti-mouse IgG to detect ER ( green ). Expressed scFv-Fc or Fc proteins were stained with Cy3-conjugated anti-human IgG ( red ). Yellow ( orange when red is stronger than green ) areas in ER/Fc Merge represent colocalized signals. Fixed cells were also treated with DAPI to stain nuclei ( blue ). Right panel , costaining of scFv-Fc or Fc proteins and Golgi apparatus. COS-7 cells transfected with scFv-Fc genes encoding 3F1, 1F12, 5A3, 1A4, 1G4, or Fc and cultured for 16 h were fixed. Fixed cells were stained with BODIPY FL C 5 -ceramide to detect Golgi apparatuses ( green ). Expressed scFv-Fc or Fc proteins were stained with Cy3-conjugated anti-human IgG ( red ). Shown are the Golgi/Fc Merge only. Fixed cells were also treated with DAPI (1:1,000 dilutions) to stain nuclei ( blue ). Dilutions used for primary and secondary antibodies were anti-PDI (1:100), FITC-conjugated AffiniPure donkey anti-mouse IgG (1:100), BODIPY FL C 5 -ceramide (1:200), and Cy3-conjugated anti-human IgG (1:500). The white bar indicates 20 μm.
    Figure Legend Snippet: Colocalization of scFv-Fc or Fc proteins with ER. Left panel , COS-7 cells transfected with scFv-Fc genes encoding 3F1, 1F12, 5A3, 1A4, 1G4, or Fc and cultured for 16 h were fixed. Fixed cells were stained with anti-PDI antibody and FITC-conjugated AffiniPure donkey anti-mouse IgG to detect ER ( green ). Expressed scFv-Fc or Fc proteins were stained with Cy3-conjugated anti-human IgG ( red ). Yellow ( orange when red is stronger than green ) areas in ER/Fc Merge represent colocalized signals. Fixed cells were also treated with DAPI to stain nuclei ( blue ). Right panel , costaining of scFv-Fc or Fc proteins and Golgi apparatus. COS-7 cells transfected with scFv-Fc genes encoding 3F1, 1F12, 5A3, 1A4, 1G4, or Fc and cultured for 16 h were fixed. Fixed cells were stained with BODIPY FL C 5 -ceramide to detect Golgi apparatuses ( green ). Expressed scFv-Fc or Fc proteins were stained with Cy3-conjugated anti-human IgG ( red ). Shown are the Golgi/Fc Merge only. Fixed cells were also treated with DAPI (1:1,000 dilutions) to stain nuclei ( blue ). Dilutions used for primary and secondary antibodies were anti-PDI (1:100), FITC-conjugated AffiniPure donkey anti-mouse IgG (1:100), BODIPY FL C 5 -ceramide (1:200), and Cy3-conjugated anti-human IgG (1:500). The white bar indicates 20 μm.

    Techniques Used: Transfection, Cell Culture, Staining

    4) Product Images from "Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1 2"

    Article Title: Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1Calcium Channel Blocker Verapamil Enhances Endoplasmic Reticulum Stress and Cell Death Induced by Proteasome Inhibition in Myeloma Cells 1 2

    Journal: Neoplasia (New York, N.Y.)

    doi:

    JK-6L cells show enhanced protein accumulation in the presence of bortezomib and verapamil. (A) Western blot analysis of detergent-soluble and detergent-insoluble fractions of JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 16 hours. Immunoblot analysiswas done using an anti-ubiquitin antibody. To detect free ubiquitin, a denser gel was used. One representative immunoblot of two independent experiments is shown. (B) Western blot analysis of total cell lysates fromJK-6L cells treated with 10 nM bortezomib and/or 70 µMverapamil for 16 hours. Immunoblot analysis was done using HRP-conjugated anti-IgG antibody. One representative immunoblot of two independent experiments is shown. (C) IgG concentration in the supernatant of JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 16 hourswas determined by ELISA. The absorbance (OD)wasmeasured using a spectrophotometer. Mean values and SDwere calculated fromhexaplicates. Data represent one of two independently done experiments. Student's t -test for unpaired heteroscedastic samples was used for statistical analysis. # P
    Figure Legend Snippet: JK-6L cells show enhanced protein accumulation in the presence of bortezomib and verapamil. (A) Western blot analysis of detergent-soluble and detergent-insoluble fractions of JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 16 hours. Immunoblot analysiswas done using an anti-ubiquitin antibody. To detect free ubiquitin, a denser gel was used. One representative immunoblot of two independent experiments is shown. (B) Western blot analysis of total cell lysates fromJK-6L cells treated with 10 nM bortezomib and/or 70 µMverapamil for 16 hours. Immunoblot analysis was done using HRP-conjugated anti-IgG antibody. One representative immunoblot of two independent experiments is shown. (C) IgG concentration in the supernatant of JK-6L cells treated with 10 nM bortezomib and/or 70 µ M verapamil for 16 hourswas determined by ELISA. The absorbance (OD)wasmeasured using a spectrophotometer. Mean values and SDwere calculated fromhexaplicates. Data represent one of two independently done experiments. Student's t -test for unpaired heteroscedastic samples was used for statistical analysis. # P

    Techniques Used: Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Spectrophotometry

    5) Product Images from "Measurements of auto-antibodies to α-synuclein in the serum and cerebral spinal fluids of patients with Parkinson’s disease"

    Article Title: Measurements of auto-antibodies to α-synuclein in the serum and cerebral spinal fluids of patients with Parkinson’s disease

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.14330

    Optimization of ELISA for α-syn AAb detection Dilute serum was applied to 384-well ELISA plates either coated with recombinant α-syn or not coated. (A) Plates were blocked with either 1% cold fish gelatin (top row) or 5% bovine serum albumin (middle row) overnight at 4 °C. Binding was detected using anti-human IgG HRP-conjugates raised in rabbit (left column), donkey (middle column), or goat (right column). (B) Plates were blocked with 1% cold fish gelatin at 4°C for either 6 days (left), 4 days (middle), or 1 day (right). Additional blocking solutions (1% bovine serum albumin, 5% milk in PBS, several commercial blocking solutions) did not demonstrate any improvement over 1% cold fish gelatin. Each data point represents three technical replicates (n=3 wells). Wells coated with α-syn denoted by closed circles. Uncoated wells denoted by closed triangles. BSA = bovine serum albumin; CFG = cold fish gelatin; HRP = horseradish peroxidase conjugate.
    Figure Legend Snippet: Optimization of ELISA for α-syn AAb detection Dilute serum was applied to 384-well ELISA plates either coated with recombinant α-syn or not coated. (A) Plates were blocked with either 1% cold fish gelatin (top row) or 5% bovine serum albumin (middle row) overnight at 4 °C. Binding was detected using anti-human IgG HRP-conjugates raised in rabbit (left column), donkey (middle column), or goat (right column). (B) Plates were blocked with 1% cold fish gelatin at 4°C for either 6 days (left), 4 days (middle), or 1 day (right). Additional blocking solutions (1% bovine serum albumin, 5% milk in PBS, several commercial blocking solutions) did not demonstrate any improvement over 1% cold fish gelatin. Each data point represents three technical replicates (n=3 wells). Wells coated with α-syn denoted by closed circles. Uncoated wells denoted by closed triangles. BSA = bovine serum albumin; CFG = cold fish gelatin; HRP = horseradish peroxidase conjugate.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Recombinant, Fluorescence In Situ Hybridization, Binding Assay, Blocking Assay

    6) Product Images from "Targeting Effector Memory T Cells with the Small Molecule Kv1.3 Blocker PAP-1 Suppresses Allergic Contact Dermatitis"

    Article Title: Targeting Effector Memory T Cells with the Small Molecule Kv1.3 Blocker PAP-1 Suppresses Allergic Contact Dermatitis

    Journal:

    doi: 10.1038/sj.jid.5700717

    Ear infiltrating cells in ACD are CD8 + , CD45RC − , and Kv1.3 +
    Figure Legend Snippet: Ear infiltrating cells in ACD are CD8 + , CD45RC − , and Kv1.3 +

    Techniques Used:

    7) Product Images from "Targeting Effector Memory T Cells with the Small Molecule Kv1.3 Blocker PAP-1 Suppresses Allergic Contact Dermatitis"

    Article Title: Targeting Effector Memory T Cells with the Small Molecule Kv1.3 Blocker PAP-1 Suppresses Allergic Contact Dermatitis

    Journal:

    doi: 10.1038/sj.jid.5700717

    Ear infiltrating cells in ACD are CD8 + , CD45RC − , and Kv1.3 +
    Figure Legend Snippet: Ear infiltrating cells in ACD are CD8 + , CD45RC − , and Kv1.3 +

    Techniques Used:

    8) Product Images from "Calreticulin Couples Calcium Release and Calcium Influx in Integrin-mediated Calcium Signaling"

    Article Title: Calreticulin Couples Calcium Release and Calcium Influx in Integrin-mediated Calcium Signaling

    Journal: Molecular Biology of the Cell

    doi:

    Association of integrin α7 with ectocalreticulin and DHPR α1 subunit in E63 cells. (A) The unfixed E63 cells (5 d old) were first immunostained with calreticulin Ab (PA3-900) and then stained with integrin α7 Ab (O26 mAb) again. Ectocalreticulin was diffusely distributed on the cell surface (a), whereas integrin α7 displayed a stress fiber-like distribution (b). In contrast, prior incubation of integrin α7 Ab (c–h) before staining of calreticulin Ab (PA3-900 [c and g] or LAR090 [e]) induced cellular localization of ectocalreticulin (c, e, and g), in which ectocalreticulin appears to be colocalized with integrin α7 (d, f, and h). In control experiments, cells were first stained with normal mouse immunoglobulin and then stained with normal rabbit immunoglobulin (i and j). Cells shown in the left panels (a, c, e, g, and i) were stained with TRITC-conjugated donkey anti-rabbit immunoglobulin, whereas those in the right panels (b, d, f, h, and j) were stained with FITC-conjugated donkey anti-mouse immunoglobulin. Bars: in f (for a–f, i, and j), 10 μm; in h (for g and j), 5 μm. (B) Cell surface proteins in 5-d cultured cells were iodinated with Na 125 I and immunoprecipitated with calreticulin Ab (PA3-900). Autoradiography of cell surface proteins (a) and imunoblot analysis (b) of cell lysates with calreticulin Ab (PA3-900) are presented. Iodinated ectocalreticulin is indicated by 125 I-CRT. (C) Cell lysates were immunoprecipitated (IP) using integrin α7 Ab (H36 mAb) and DHPR α1 Ab and then immunoblotted (IB) with calreticulin Ab: PA3-900 (left panel) and LAR090 (right panel). (D) After stripping, the same membrane was reprobed with DHPR α1 Ab. Note that integrin α7 was associated with both calreticulin and DHPR α1.
    Figure Legend Snippet: Association of integrin α7 with ectocalreticulin and DHPR α1 subunit in E63 cells. (A) The unfixed E63 cells (5 d old) were first immunostained with calreticulin Ab (PA3-900) and then stained with integrin α7 Ab (O26 mAb) again. Ectocalreticulin was diffusely distributed on the cell surface (a), whereas integrin α7 displayed a stress fiber-like distribution (b). In contrast, prior incubation of integrin α7 Ab (c–h) before staining of calreticulin Ab (PA3-900 [c and g] or LAR090 [e]) induced cellular localization of ectocalreticulin (c, e, and g), in which ectocalreticulin appears to be colocalized with integrin α7 (d, f, and h). In control experiments, cells were first stained with normal mouse immunoglobulin and then stained with normal rabbit immunoglobulin (i and j). Cells shown in the left panels (a, c, e, g, and i) were stained with TRITC-conjugated donkey anti-rabbit immunoglobulin, whereas those in the right panels (b, d, f, h, and j) were stained with FITC-conjugated donkey anti-mouse immunoglobulin. Bars: in f (for a–f, i, and j), 10 μm; in h (for g and j), 5 μm. (B) Cell surface proteins in 5-d cultured cells were iodinated with Na 125 I and immunoprecipitated with calreticulin Ab (PA3-900). Autoradiography of cell surface proteins (a) and imunoblot analysis (b) of cell lysates with calreticulin Ab (PA3-900) are presented. Iodinated ectocalreticulin is indicated by 125 I-CRT. (C) Cell lysates were immunoprecipitated (IP) using integrin α7 Ab (H36 mAb) and DHPR α1 Ab and then immunoblotted (IB) with calreticulin Ab: PA3-900 (left panel) and LAR090 (right panel). (D) After stripping, the same membrane was reprobed with DHPR α1 Ab. Note that integrin α7 was associated with both calreticulin and DHPR α1.

    Techniques Used: Staining, Incubation, Cell Culture, Immunoprecipitation, Autoradiography, Stripping Membranes

    9) Product Images from "Keeping the Vimentin Network under Control: Cell-Matrix Adhesion-associated Plectin 1f Affects Cell Shape and Polarity of Fibroblasts"

    Article Title: Keeping the Vimentin Network under Control: Cell-Matrix Adhesion-associated Plectin 1f Affects Cell Shape and Polarity of Fibroblasts

    Journal: Molecular Biology of the Cell

    doi: 10.1091/mbc.E10-02-0094

    First exon-encoded sequence of P1f controls FA-targeting and actin-binding. (A) Schematic representation of P1f-8's domain composition and first exon-encoded isoform-specific amino acid sequence. Arrowhead, tyrosine residue at position 20. (B and C) Fluorescence microscopy of immortal (p53 −/− ) plectin-deficient fibroblasts transfected with cDNA constructs encoding EGFP-tagged plectin fragments as indicated. Scale bars, 10 μm. Note, only the wild-type P1f-8-EGFP fragment and the P1f-8F-EGFP mutant protein are targeted to FAs. (D) Cosedimentation of P1f mutant proteins with F-actin. Recombinant plectin fragments indicated were incubated with preassembled F-actin. F-actin and proteins bound were sedimented by centrifugation, and equal amounts of supernatant (S) and pellet (P) fractions were subjected to SDS-PAGE; separated proteins were visualized by Comassie Brilliant Blue staining. (E) Densitometric quantitation of gel bands. Data shown represent mean values (±SEM) of three independent experiments, including the one shown in D.
    Figure Legend Snippet: First exon-encoded sequence of P1f controls FA-targeting and actin-binding. (A) Schematic representation of P1f-8's domain composition and first exon-encoded isoform-specific amino acid sequence. Arrowhead, tyrosine residue at position 20. (B and C) Fluorescence microscopy of immortal (p53 −/− ) plectin-deficient fibroblasts transfected with cDNA constructs encoding EGFP-tagged plectin fragments as indicated. Scale bars, 10 μm. Note, only the wild-type P1f-8-EGFP fragment and the P1f-8F-EGFP mutant protein are targeted to FAs. (D) Cosedimentation of P1f mutant proteins with F-actin. Recombinant plectin fragments indicated were incubated with preassembled F-actin. F-actin and proteins bound were sedimented by centrifugation, and equal amounts of supernatant (S) and pellet (P) fractions were subjected to SDS-PAGE; separated proteins were visualized by Comassie Brilliant Blue staining. (E) Densitometric quantitation of gel bands. Data shown represent mean values (±SEM) of three independent experiments, including the one shown in D.

    Techniques Used: Sequencing, Binding Assay, Fluorescence, Microscopy, Transfection, Construct, Mutagenesis, Recombinant, Incubation, Centrifugation, SDS Page, Staining, Quantitation Assay

    P1f-induced formation of vimentin filament intermediates and their association with different plectin isoforms. (A) Immunofluorescence microscopy (vimentin) after forced expression of full-length P1f-EGFP in immortal plectin −/− fibroblasts. A transfected cell (green) and a nontransfected cell (red only) are shown side by side. The boxed area corresponds to the cropped and magnified view shown in ROI. Arrows, vimentin filament intermediates colocalizing with P1f; arrowheads, vimentin filament intermediates bare of P1f. Note, that the nontransfected cell still exhibits wavy vimentin bundles extending to the cell periphery typical of plectin −/− fibroblasts. Scale bars, 20 μm, and (ROI) 5 μm. (B) Immunoblotting of cell lysate from primary mouse fibroblasts using plectin isoform-specific (1f, 1c, and 1) and anti-pan plectin (no. 9) antibodies. (C) Triple immunofluorescence microscopy (P1c, talin, and vimentin) of primary plectin +/+ fibroblasts. The bottom panels display cropped and magnified views of the boxed area in the top panel. Arrowheads, filament intermediates that are positive for P1c, but are not associated with FAs; arrows, P1c-decorated filament intermediates colocalizing with FAs. Scale bars, 20 μm, and (ROI) 5 μm. (D and E) Statistical evaluation of P1f and P1c distribution with vimentin filament intermediates at FAs. Data shown represent mean values (± the SEM) from randomly chosen cells (n = 6); on average, 150 FAs were counted per cell. *p
    Figure Legend Snippet: P1f-induced formation of vimentin filament intermediates and their association with different plectin isoforms. (A) Immunofluorescence microscopy (vimentin) after forced expression of full-length P1f-EGFP in immortal plectin −/− fibroblasts. A transfected cell (green) and a nontransfected cell (red only) are shown side by side. The boxed area corresponds to the cropped and magnified view shown in ROI. Arrows, vimentin filament intermediates colocalizing with P1f; arrowheads, vimentin filament intermediates bare of P1f. Note, that the nontransfected cell still exhibits wavy vimentin bundles extending to the cell periphery typical of plectin −/− fibroblasts. Scale bars, 20 μm, and (ROI) 5 μm. (B) Immunoblotting of cell lysate from primary mouse fibroblasts using plectin isoform-specific (1f, 1c, and 1) and anti-pan plectin (no. 9) antibodies. (C) Triple immunofluorescence microscopy (P1c, talin, and vimentin) of primary plectin +/+ fibroblasts. The bottom panels display cropped and magnified views of the boxed area in the top panel. Arrowheads, filament intermediates that are positive for P1c, but are not associated with FAs; arrows, P1c-decorated filament intermediates colocalizing with FAs. Scale bars, 20 μm, and (ROI) 5 μm. (D and E) Statistical evaluation of P1f and P1c distribution with vimentin filament intermediates at FAs. Data shown represent mean values (± the SEM) from randomly chosen cells (n = 6); on average, 150 FAs were counted per cell. *p

    Techniques Used: Immunofluorescence, Microscopy, Expressing, Transfection

    10) Product Images from "Factors Determining the Risk of Inadvertent Retroviral Transduction of Male Germ Cells After In Utero Gene Transfer in Sheep"

    Article Title: Factors Determining the Risk of Inadvertent Retroviral Transduction of Male Germ Cells After In Utero Gene Transfer in Sheep

    Journal: Human Gene Therapy

    doi: 10.1089/hum.2007.120

    SSEA-4 is a marker of GCs compartmentalized within fetal sex cords. Representative photomicrographs of medium-power magnification (× 40 objective) showing distribution of SSEA-4-labeled GCs within fetal testes. Robustly labeled GCs were
    Figure Legend Snippet: SSEA-4 is a marker of GCs compartmentalized within fetal sex cords. Representative photomicrographs of medium-power magnification (× 40 objective) showing distribution of SSEA-4-labeled GCs within fetal testes. Robustly labeled GCs were

    Techniques Used: Marker, Labeling

    11) Product Images from "Incorporating B cell activating factor (BAFF) into the membrane of rabies virus (RABV) particles improves the speed and magnitude of vaccine-induced antibody responses"

    Article Title: Incorporating B cell activating factor (BAFF) into the membrane of rabies virus (RABV) particles improves the speed and magnitude of vaccine-induced antibody responses

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0007800

    100-fold less RABV-ED51-mBAFF is needed to induce comparable immunity in mice immunized with RABV. C57BL/6J mice were immunized i.m. with 10 3 or 10 5 ffu/mouse of RABV or RABV-ED51-mBAFF, or PBS as a negative control. Blood was collected on days 5, 7 and 10 post-immunization as a source of serum. Four three-fold serial dilutions of sera were analyzed by ELISA to determine anti-RABV G IgM (A and B) or IgG (C and D) antibody titers and presented as OD 490 of the reciprocal serial dilution. For comparison, sera from PBS-immunized mice were tested in parallel. Statistical difference in antibody titers by ELISA between RABV- and RABV-ED51-mBAFF-immunized mice was determined using an unpaired, two-tailed t test and data is presented as the mean ± SEM. *p≤0.05; **p ≤ 0.01; ***p≤0.001; ****p≤0.0001; N = 10 mice/group). (ffu = focus forming units; OD = optical density).
    Figure Legend Snippet: 100-fold less RABV-ED51-mBAFF is needed to induce comparable immunity in mice immunized with RABV. C57BL/6J mice were immunized i.m. with 10 3 or 10 5 ffu/mouse of RABV or RABV-ED51-mBAFF, or PBS as a negative control. Blood was collected on days 5, 7 and 10 post-immunization as a source of serum. Four three-fold serial dilutions of sera were analyzed by ELISA to determine anti-RABV G IgM (A and B) or IgG (C and D) antibody titers and presented as OD 490 of the reciprocal serial dilution. For comparison, sera from PBS-immunized mice were tested in parallel. Statistical difference in antibody titers by ELISA between RABV- and RABV-ED51-mBAFF-immunized mice was determined using an unpaired, two-tailed t test and data is presented as the mean ± SEM. *p≤0.05; **p ≤ 0.01; ***p≤0.001; ****p≤0.0001; N = 10 mice/group). (ffu = focus forming units; OD = optical density).

    Techniques Used: Mouse Assay, Negative Control, Enzyme-linked Immunosorbent Assay, Serial Dilution, Two Tailed Test

    12) Product Images from "Enzyme-Linked Immunosorbent Assay for Detection of Filovirus Species-Specific Antibodies ▿"

    Article Title: Enzyme-Linked Immunosorbent Assay for Detection of Filovirus Species-Specific Antibodies ▿

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.00170-10

    IgG and IgM antibodies detected in human samples. OD values for specific IgG (a) and IgM (b) antibodies in the patient sera are shown. Sera from 21 individuals were analyzed at 1:1,000 dilutions. Naïve human sera (1:100 dilution) were used as a negative control. Each bar represents the average and standard deviation of data from independent experiments. Asterisks indicate statistically significant differences in OD values between the homologous antigen and all other antigens ( P
    Figure Legend Snippet: IgG and IgM antibodies detected in human samples. OD values for specific IgG (a) and IgM (b) antibodies in the patient sera are shown. Sera from 21 individuals were analyzed at 1:1,000 dilutions. Naïve human sera (1:100 dilution) were used as a negative control. Each bar represents the average and standard deviation of data from independent experiments. Asterisks indicate statistically significant differences in OD values between the homologous antigen and all other antigens ( P

    Techniques Used: Negative Control, Standard Deviation

    13) Product Images from "Dynamin is involved in human epithelial cell vacuolation caused by the Helicobacter pylori-produced cytotoxin VacA"

    Article Title: Dynamin is involved in human epithelial cell vacuolation caused by the Helicobacter pylori-produced cytotoxin VacA

    Journal: Journal of Clinical Investigation

    doi:

    Intracellular localization of clathrin in VacA-treated and nontreated HeLa cells. Naive HeLa cells ( a ) and VacA-treated HeLa cells ( c ) were fixed, stained with an anti-clathrin antibody, and visualized with FITC-conjugated anti-mouse IgG. ( b and d ) Phase-contrast images of a and c , respectively. VacA-treatment did not affect intracellular distribution of clathrin in HeLa cells. Bars, 10 μm. ×400.
    Figure Legend Snippet: Intracellular localization of clathrin in VacA-treated and nontreated HeLa cells. Naive HeLa cells ( a ) and VacA-treated HeLa cells ( c ) were fixed, stained with an anti-clathrin antibody, and visualized with FITC-conjugated anti-mouse IgG. ( b and d ) Phase-contrast images of a and c , respectively. VacA-treatment did not affect intracellular distribution of clathrin in HeLa cells. Bars, 10 μm. ×400.

    Techniques Used: Staining

    14) Product Images from "A Spontaneous Deletion within the Desmoglein 3 Extracellular Domain of Mice Results in Hypomorphic Protein Expression, Immunodeficiency, and a Wasting Disease Phenotype"

    Article Title: A Spontaneous Deletion within the Desmoglein 3 Extracellular Domain of Mice Results in Hypomorphic Protein Expression, Immunodeficiency, and a Wasting Disease Phenotype

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2014.10.025

    A desmoglein 3 ( Dsg3 ) gene segment deletion in squeaky ( sqk ) mice. A: Dsg3 gene organization in wild-type (WT) mice. A ruler indicates the gene position on chromosome 18, with each horizontal bar equal to 2 Kbp. The Dsg3 + gene contains 16 exons depicted
    Figure Legend Snippet: A desmoglein 3 ( Dsg3 ) gene segment deletion in squeaky ( sqk ) mice. A: Dsg3 gene organization in wild-type (WT) mice. A ruler indicates the gene position on chromosome 18, with each horizontal bar equal to 2 Kbp. The Dsg3 + gene contains 16 exons depicted

    Techniques Used: Mouse Assay

    Dsg3 sqk/sqk and malnourished wild-type (WT) mice share similar phenotypes. A: Dsg3 sqk/sqk mice on solid-food diets have reduced body weights, but gain weight when the diet is supplemented with Ensure Plus starting at birth. Weights of WT (black triangles),
    Figure Legend Snippet: Dsg3 sqk/sqk and malnourished wild-type (WT) mice share similar phenotypes. A: Dsg3 sqk/sqk mice on solid-food diets have reduced body weights, but gain weight when the diet is supplemented with Ensure Plus starting at birth. Weights of WT (black triangles),

    Techniques Used: Mouse Assay

    Trachea, snout, and tongue tissue sections from Dsg3 sqk/sqk and Dsg3 +/+ littermates. A: Trachea cross sections obtained from 6-month-old mice of each genotype show a thickening of the pseudostratified ciliated columnar epithelium in Dsg3 sqk/sqk mice.
    Figure Legend Snippet: Trachea, snout, and tongue tissue sections from Dsg3 sqk/sqk and Dsg3 +/+ littermates. A: Trachea cross sections obtained from 6-month-old mice of each genotype show a thickening of the pseudostratified ciliated columnar epithelium in Dsg3 sqk/sqk mice.

    Techniques Used: Mouse Assay

    Dsg3 mRNA and protein expression in Dsg3 +/+ and Dsg3 sqk/sqk mice. A: Dsg3 sqk/sqk , Dsg3 sqk/+ , and Dsg3 +/+ mice express Dgs3 transcripts equally. The diagram indicates the positions of the Dsg3 cDNA-specific primer pair relative to gene structure. The reverse
    Figure Legend Snippet: Dsg3 mRNA and protein expression in Dsg3 +/+ and Dsg3 sqk/sqk mice. A: Dsg3 sqk/sqk , Dsg3 sqk/+ , and Dsg3 +/+ mice express Dgs3 transcripts equally. The diagram indicates the positions of the Dsg3 cDNA-specific primer pair relative to gene structure. The reverse

    Techniques Used: Expressing, Mouse Assay

    15) Product Images from "Isolation of human monoclonal antibodies from peripheral blood B cells"

    Article Title: Isolation of human monoclonal antibodies from peripheral blood B cells

    Journal: Nature protocols

    doi: 10.1038/nprot.2013.117

    Gating strategy for a memory B cell population. ( a ) Gating of CD19 + IgM − IgA − IgD − memory B cells from a sample of PBMCs collected in a patient with HIV infection. ( b ) Purity of CD19 + IgM − IgA − IgD − B cells after
    Figure Legend Snippet: Gating strategy for a memory B cell population. ( a ) Gating of CD19 + IgM − IgA − IgD − memory B cells from a sample of PBMCs collected in a patient with HIV infection. ( b ) Purity of CD19 + IgM − IgA − IgD − B cells after

    Techniques Used: Infection

    16) Product Images from "Factors Determining the Risk of Inadvertent Retroviral Transduction of Male Germ Cells After In Utero Gene Transfer in Sheep"

    Article Title: Factors Determining the Risk of Inadvertent Retroviral Transduction of Male Germ Cells After In Utero Gene Transfer in Sheep

    Journal: Human Gene Therapy

    doi: 10.1089/hum.2007.120

    SSEA-3 is a marker of GC transition from migratory state to sex cord compartmentalization within the fetal testes. Photomicrographs taken at medium-power magnification (× 40 objective) show the distribution of SSEA-3-labeled GCs. The anti-SSEA-3
    Figure Legend Snippet: SSEA-3 is a marker of GC transition from migratory state to sex cord compartmentalization within the fetal testes. Photomicrographs taken at medium-power magnification (× 40 objective) show the distribution of SSEA-3-labeled GCs. The anti-SSEA-3

    Techniques Used: Marker, Labeling

    Distribution of SSEA-3-labeled GCs of fetal testes on gestational day 68. Representative low-power (×10 objective) photomicrograph shows SSEA-3-labeled GCs within the fetal testis. Note migrating GCs within the internal rete testes (R) and the
    Figure Legend Snippet: Distribution of SSEA-3-labeled GCs of fetal testes on gestational day 68. Representative low-power (×10 objective) photomicrograph shows SSEA-3-labeled GCs within the fetal testis. Note migrating GCs within the internal rete testes (R) and the

    Techniques Used: Labeling

    17) Product Images from "Targeting Effector Memory T Cells with the Small Molecule Kv1.3 Blocker PAP-1 Suppresses Allergic Contact Dermatitis"

    Article Title: Targeting Effector Memory T Cells with the Small Molecule Kv1.3 Blocker PAP-1 Suppresses Allergic Contact Dermatitis

    Journal:

    doi: 10.1038/sj.jid.5700717

    PAP-1 suppresses oxazolone-induced inflammation by reducing CD8+ T cells infiltration and production of the inflammatory cytokines IFN- γ and IL-17 but not TNF- α
    Figure Legend Snippet: PAP-1 suppresses oxazolone-induced inflammation by reducing CD8+ T cells infiltration and production of the inflammatory cytokines IFN- γ and IL-17 but not TNF- α

    Techniques Used:

    Ear infiltrating cells in ACD are CD8 + , CD45RC − , and Kv1.3 +
    Figure Legend Snippet: Ear infiltrating cells in ACD are CD8 + , CD45RC − , and Kv1.3 +

    Techniques Used:

    18) Product Images from "Cell Surface Proteome of Dental Pulp Stem Cells Identified by Label-Free Mass Spectrometry"

    Article Title: Cell Surface Proteome of Dental Pulp Stem Cells Identified by Label-Free Mass Spectrometry

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0159824

    Differential expression of MS-based identified surface proteins by DPSCs cultured in different media. RT-PCR ( A and B) and immunoblotting ( C and D) analyses revealed the expression level of selected MS-identified cell surface markers of DPSCs cultured either in basic expansion (BE) or standard (S) media. Samples from 5 independent donors from both culture conditions were analyzed (donors 1–5 and 6–10, respectively). MS data were derived from cells obtained from donors 1 and 6 (blue frame). The markers at the top were found by MS to be predominantly expressed on cells growing in BE medium, whereas the others were mainly found after culture in S medium. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control. A. M: 100 base pair DNA Ladder, c: Negative control (i.e. without cDNA template) C. The positions of prestained molecular mass markers are indicated on the left. B and D. Expression level of specific cell surface markers from donors 1 and 6 was quantified. Data are the mean of two to three independent experiments using distinct mRNA and protein preparations; bars indicate the variation of the individual values from the mean. The expression level was arbitrarily set to 100% for the marker predominantly identified under a specific culture condition. Note that the expression of protein markers enriched in DPSCs cultured in S medium fluctuates between donors as illustrated by CADH2 and MFGM (see also S8 and S9 Figs).
    Figure Legend Snippet: Differential expression of MS-based identified surface proteins by DPSCs cultured in different media. RT-PCR ( A and B) and immunoblotting ( C and D) analyses revealed the expression level of selected MS-identified cell surface markers of DPSCs cultured either in basic expansion (BE) or standard (S) media. Samples from 5 independent donors from both culture conditions were analyzed (donors 1–5 and 6–10, respectively). MS data were derived from cells obtained from donors 1 and 6 (blue frame). The markers at the top were found by MS to be predominantly expressed on cells growing in BE medium, whereas the others were mainly found after culture in S medium. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as loading control. A. M: 100 base pair DNA Ladder, c: Negative control (i.e. without cDNA template) C. The positions of prestained molecular mass markers are indicated on the left. B and D. Expression level of specific cell surface markers from donors 1 and 6 was quantified. Data are the mean of two to three independent experiments using distinct mRNA and protein preparations; bars indicate the variation of the individual values from the mean. The expression level was arbitrarily set to 100% for the marker predominantly identified under a specific culture condition. Note that the expression of protein markers enriched in DPSCs cultured in S medium fluctuates between donors as illustrated by CADH2 and MFGM (see also S8 and S9 Figs).

    Techniques Used: Expressing, Mass Spectrometry, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Negative Control, Marker

    19) Product Images from "Carbonic anhydrase II-positive pancreatic cells are progenitors for both endocrine and exocrine pancreas after birth"

    Article Title: Carbonic anhydrase II-positive pancreatic cells are progenitors for both endocrine and exocrine pancreas after birth

    Journal:

    doi: 10.1073/pnas.0805803105

    Characterization of transgene expression. ( A–C ) At birth β-galactosidase immunostaining (green) was seen in ducts (d) and nerve ganglion (g) but not in insulin-positive beta cells (red; A and B ), glucagon-positive cells (red; C ) or acini
    Figure Legend Snippet: Characterization of transgene expression. ( A–C ) At birth β-galactosidase immunostaining (green) was seen in ducts (d) and nerve ganglion (g) but not in insulin-positive beta cells (red; A and B ), glucagon-positive cells (red; C ) or acini

    Techniques Used: Expressing, Immunostaining

    20) Product Images from "Cytokeratin 8 Is an Epithelial Cell Receptor for Pet, a Cytotoxic Serine Protease Autotransporter of Enterobacteriaceae"

    Article Title: Cytokeratin 8 Is an Epithelial Cell Receptor for Pet, a Cytotoxic Serine Protease Autotransporter of Enterobacteriaceae

    Journal: mBio

    doi: 10.1128/mBio.00838-13

    Pet bound to CK8 undergoes clathrin-mediated endocytosis. HEp-2 cells were treated with Pet (37 µg/ml) for 1, 2, 4, or 8 min. Cells were then washed, fixed, and permeabilized. Clathrin was detected using a polyclonal anti-clathrin antibody followed by a secondary anti-goat IgG antibody coupled to fluorescein (green), CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary antibody anti-mouse IgG coupled to rhodamine (red), and Pet was detected using a rabbit anti-Pet polyclonal antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Pet bound to CK8 undergoes clathrin-mediated endocytosis. HEp-2 cells were treated with Pet (37 µg/ml) for 1, 2, 4, or 8 min. Cells were then washed, fixed, and permeabilized. Clathrin was detected using a polyclonal anti-clathrin antibody followed by a secondary anti-goat IgG antibody coupled to fluorescein (green), CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary antibody anti-mouse IgG coupled to rhodamine (red), and Pet was detected using a rabbit anti-Pet polyclonal antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Confocal Microscopy

    Kidney cells transfected with ck8 are susceptible to cell damage caused by Pet. At 72 h posttransfection, MDCK cells transfected with ck8-gfp were treated with Pet (37 µg/ml) for 6 h. Untransfected and transfected cells were fixed, permeabilized, and stained with rhodamine-phalloidin for F actin (red). Pet was detected using a rabbit polyclonal anti-Pet antibody followed by anti-rabbit IgG secondary antibodies labeled with Cy5 (blue). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with fluorescein (green). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Kidney cells transfected with ck8 are susceptible to cell damage caused by Pet. At 72 h posttransfection, MDCK cells transfected with ck8-gfp were treated with Pet (37 µg/ml) for 6 h. Untransfected and transfected cells were fixed, permeabilized, and stained with rhodamine-phalloidin for F actin (red). Pet was detected using a rabbit polyclonal anti-Pet antibody followed by anti-rabbit IgG secondary antibodies labeled with Cy5 (blue). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with fluorescein (green). Slides were observed by confocal microscopy.

    Techniques Used: Transfection, Positron Emission Tomography, Staining, Labeling, Confocal Microscopy

    Pet and CK8 colocalize as a ligand-receptor complex in early endosomes of epithelial cells. HEp-2 cells were treated overnight with the CellLight early endosomes-RFP BacMam 2.0 reagent to mark Rab5b, a resident protein of early endosomes (red). Cells treated with Pet for 7, 8, 9, or 10 min were then washed, fixed, and permeabilized. CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary anti-mouse IgG antibody coupled to fluorescein (green), and Pet was detected using a rabbit polyclonal anti-Pet antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Pet and CK8 colocalize as a ligand-receptor complex in early endosomes of epithelial cells. HEp-2 cells were treated overnight with the CellLight early endosomes-RFP BacMam 2.0 reagent to mark Rab5b, a resident protein of early endosomes (red). Cells treated with Pet for 7, 8, 9, or 10 min were then washed, fixed, and permeabilized. CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary anti-mouse IgG antibody coupled to fluorescein (green), and Pet was detected using a rabbit polyclonal anti-Pet antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Confocal Microscopy

    Knockdown of CK8 prevents cell damage induced by Pet. (A) Knockdown of CK8 by siRNA. HEp-2 cells were silenced using siRNA for ck8 . Cells were lysed and analyzed by Western blotting using anti-CK8 monoclonal antibodies in comparison to normal cells. Results were analyzed by densitometry and plotted as the ratio of CK8 to actin. (B) Effect of Pet on CK8 knockdown cells. HEp-2 cells silenced using siRNA for ck8 were then treated with Pet (37 µg/ml) for 3 h, fixed, permeabilized, and stained with rhodaminated phalloidin (red). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Knockdown of CK8 prevents cell damage induced by Pet. (A) Knockdown of CK8 by siRNA. HEp-2 cells were silenced using siRNA for ck8 . Cells were lysed and analyzed by Western blotting using anti-CK8 monoclonal antibodies in comparison to normal cells. Results were analyzed by densitometry and plotted as the ratio of CK8 to actin. (B) Effect of Pet on CK8 knockdown cells. HEp-2 cells silenced using siRNA for ck8 were then treated with Pet (37 µg/ml) for 3 h, fixed, permeabilized, and stained with rhodaminated phalloidin (red). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Western Blot, Staining, Labeling, Confocal Microscopy

    21) Product Images from "Cytokeratin 8 Is an Epithelial Cell Receptor for Pet, a Cytotoxic Serine Protease Autotransporter of Enterobacteriaceae"

    Article Title: Cytokeratin 8 Is an Epithelial Cell Receptor for Pet, a Cytotoxic Serine Protease Autotransporter of Enterobacteriaceae

    Journal: mBio

    doi: 10.1128/mBio.00838-13

    Pet bound to CK8 undergoes clathrin-mediated endocytosis. HEp-2 cells were treated with Pet (37 µg/ml) for 1, 2, 4, or 8 min. Cells were then washed, fixed, and permeabilized. Clathrin was detected using a polyclonal anti-clathrin antibody followed by a secondary anti-goat IgG antibody coupled to fluorescein (green), CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary antibody anti-mouse IgG coupled to rhodamine (red), and Pet was detected using a rabbit anti-Pet polyclonal antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Pet bound to CK8 undergoes clathrin-mediated endocytosis. HEp-2 cells were treated with Pet (37 µg/ml) for 1, 2, 4, or 8 min. Cells were then washed, fixed, and permeabilized. Clathrin was detected using a polyclonal anti-clathrin antibody followed by a secondary anti-goat IgG antibody coupled to fluorescein (green), CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary antibody anti-mouse IgG coupled to rhodamine (red), and Pet was detected using a rabbit anti-Pet polyclonal antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Confocal Microscopy

    Kidney cells transfected with ck8 are susceptible to cell damage caused by Pet. At 72 h posttransfection, MDCK cells transfected with ck8-gfp were treated with Pet (37 µg/ml) for 6 h. Untransfected and transfected cells were fixed, permeabilized, and stained with rhodamine-phalloidin for F actin (red). Pet was detected using a rabbit polyclonal anti-Pet antibody followed by anti-rabbit IgG secondary antibodies labeled with Cy5 (blue). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with fluorescein (green). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Kidney cells transfected with ck8 are susceptible to cell damage caused by Pet. At 72 h posttransfection, MDCK cells transfected with ck8-gfp were treated with Pet (37 µg/ml) for 6 h. Untransfected and transfected cells were fixed, permeabilized, and stained with rhodamine-phalloidin for F actin (red). Pet was detected using a rabbit polyclonal anti-Pet antibody followed by anti-rabbit IgG secondary antibodies labeled with Cy5 (blue). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with fluorescein (green). Slides were observed by confocal microscopy.

    Techniques Used: Transfection, Positron Emission Tomography, Staining, Labeling, Confocal Microscopy

    Pet and CK8 colocalize as a ligand-receptor complex in early endosomes of epithelial cells. HEp-2 cells were treated overnight with the CellLight early endosomes-RFP BacMam 2.0 reagent to mark Rab5b, a resident protein of early endosomes (red). Cells treated with Pet for 7, 8, 9, or 10 min were then washed, fixed, and permeabilized. CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary anti-mouse IgG antibody coupled to fluorescein (green), and Pet was detected using a rabbit polyclonal anti-Pet antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Pet and CK8 colocalize as a ligand-receptor complex in early endosomes of epithelial cells. HEp-2 cells were treated overnight with the CellLight early endosomes-RFP BacMam 2.0 reagent to mark Rab5b, a resident protein of early endosomes (red). Cells treated with Pet for 7, 8, 9, or 10 min were then washed, fixed, and permeabilized. CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary anti-mouse IgG antibody coupled to fluorescein (green), and Pet was detected using a rabbit polyclonal anti-Pet antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Confocal Microscopy

    Knockdown of CK8 prevents cell damage induced by Pet. (A) Knockdown of CK8 by siRNA. HEp-2 cells were silenced using siRNA for ck8 . Cells were lysed and analyzed by Western blotting using anti-CK8 monoclonal antibodies in comparison to normal cells. Results were analyzed by densitometry and plotted as the ratio of CK8 to actin. (B) Effect of Pet on CK8 knockdown cells. HEp-2 cells silenced using siRNA for ck8 were then treated with Pet (37 µg/ml) for 3 h, fixed, permeabilized, and stained with rhodaminated phalloidin (red). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Knockdown of CK8 prevents cell damage induced by Pet. (A) Knockdown of CK8 by siRNA. HEp-2 cells were silenced using siRNA for ck8 . Cells were lysed and analyzed by Western blotting using anti-CK8 monoclonal antibodies in comparison to normal cells. Results were analyzed by densitometry and plotted as the ratio of CK8 to actin. (B) Effect of Pet on CK8 knockdown cells. HEp-2 cells silenced using siRNA for ck8 were then treated with Pet (37 µg/ml) for 3 h, fixed, permeabilized, and stained with rhodaminated phalloidin (red). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Western Blot, Staining, Labeling, Confocal Microscopy

    22) Product Images from "Cytokeratin 8 Is an Epithelial Cell Receptor for Pet, a Cytotoxic Serine Protease Autotransporter of Enterobacteriaceae"

    Article Title: Cytokeratin 8 Is an Epithelial Cell Receptor for Pet, a Cytotoxic Serine Protease Autotransporter of Enterobacteriaceae

    Journal: mBio

    doi: 10.1128/mBio.00838-13

    Pet bound to CK8 undergoes clathrin-mediated endocytosis. HEp-2 cells were treated with Pet (37 µg/ml) for 1, 2, 4, or 8 min. Cells were then washed, fixed, and permeabilized. Clathrin was detected using a polyclonal anti-clathrin antibody followed by a secondary anti-goat IgG antibody coupled to fluorescein (green), CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary antibody anti-mouse IgG coupled to rhodamine (red), and Pet was detected using a rabbit anti-Pet polyclonal antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Pet bound to CK8 undergoes clathrin-mediated endocytosis. HEp-2 cells were treated with Pet (37 µg/ml) for 1, 2, 4, or 8 min. Cells were then washed, fixed, and permeabilized. Clathrin was detected using a polyclonal anti-clathrin antibody followed by a secondary anti-goat IgG antibody coupled to fluorescein (green), CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary antibody anti-mouse IgG coupled to rhodamine (red), and Pet was detected using a rabbit anti-Pet polyclonal antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Confocal Microscopy

    Kidney cells transfected with ck8 are susceptible to cell damage caused by Pet. At 72 h posttransfection, MDCK cells transfected with ck8-gfp were treated with Pet (37 µg/ml) for 6 h. Untransfected and transfected cells were fixed, permeabilized, and stained with rhodamine-phalloidin for F actin (red). Pet was detected using a rabbit polyclonal anti-Pet antibody followed by anti-rabbit IgG secondary antibodies labeled with Cy5 (blue). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with fluorescein (green). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Kidney cells transfected with ck8 are susceptible to cell damage caused by Pet. At 72 h posttransfection, MDCK cells transfected with ck8-gfp were treated with Pet (37 µg/ml) for 6 h. Untransfected and transfected cells were fixed, permeabilized, and stained with rhodamine-phalloidin for F actin (red). Pet was detected using a rabbit polyclonal anti-Pet antibody followed by anti-rabbit IgG secondary antibodies labeled with Cy5 (blue). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with fluorescein (green). Slides were observed by confocal microscopy.

    Techniques Used: Transfection, Positron Emission Tomography, Staining, Labeling, Confocal Microscopy

    Pet and CK8 colocalize as a ligand-receptor complex in early endosomes of epithelial cells. HEp-2 cells were treated overnight with the CellLight early endosomes-RFP BacMam 2.0 reagent to mark Rab5b, a resident protein of early endosomes (red). Cells treated with Pet for 7, 8, 9, or 10 min were then washed, fixed, and permeabilized. CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary anti-mouse IgG antibody coupled to fluorescein (green), and Pet was detected using a rabbit polyclonal anti-Pet antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Pet and CK8 colocalize as a ligand-receptor complex in early endosomes of epithelial cells. HEp-2 cells were treated overnight with the CellLight early endosomes-RFP BacMam 2.0 reagent to mark Rab5b, a resident protein of early endosomes (red). Cells treated with Pet for 7, 8, 9, or 10 min were then washed, fixed, and permeabilized. CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary anti-mouse IgG antibody coupled to fluorescein (green), and Pet was detected using a rabbit polyclonal anti-Pet antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Confocal Microscopy

    Knockdown of CK8 prevents cell damage induced by Pet. (A) Knockdown of CK8 by siRNA. HEp-2 cells were silenced using siRNA for ck8 . Cells were lysed and analyzed by Western blotting using anti-CK8 monoclonal antibodies in comparison to normal cells. Results were analyzed by densitometry and plotted as the ratio of CK8 to actin. (B) Effect of Pet on CK8 knockdown cells. HEp-2 cells silenced using siRNA for ck8 were then treated with Pet (37 µg/ml) for 3 h, fixed, permeabilized, and stained with rhodaminated phalloidin (red). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Knockdown of CK8 prevents cell damage induced by Pet. (A) Knockdown of CK8 by siRNA. HEp-2 cells were silenced using siRNA for ck8 . Cells were lysed and analyzed by Western blotting using anti-CK8 monoclonal antibodies in comparison to normal cells. Results were analyzed by densitometry and plotted as the ratio of CK8 to actin. (B) Effect of Pet on CK8 knockdown cells. HEp-2 cells silenced using siRNA for ck8 were then treated with Pet (37 µg/ml) for 3 h, fixed, permeabilized, and stained with rhodaminated phalloidin (red). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Western Blot, Staining, Labeling, Confocal Microscopy

    23) Product Images from "Cytokeratin 8 Is an Epithelial Cell Receptor for Pet, a Cytotoxic Serine Protease Autotransporter of Enterobacteriaceae"

    Article Title: Cytokeratin 8 Is an Epithelial Cell Receptor for Pet, a Cytotoxic Serine Protease Autotransporter of Enterobacteriaceae

    Journal: mBio

    doi: 10.1128/mBio.00838-13

    Pet bound to CK8 undergoes clathrin-mediated endocytosis. HEp-2 cells were treated with Pet (37 µg/ml) for 1, 2, 4, or 8 min. Cells were then washed, fixed, and permeabilized. Clathrin was detected using a polyclonal anti-clathrin antibody followed by a secondary anti-goat IgG antibody coupled to fluorescein (green), CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary antibody anti-mouse IgG coupled to rhodamine (red), and Pet was detected using a rabbit anti-Pet polyclonal antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Pet bound to CK8 undergoes clathrin-mediated endocytosis. HEp-2 cells were treated with Pet (37 µg/ml) for 1, 2, 4, or 8 min. Cells were then washed, fixed, and permeabilized. Clathrin was detected using a polyclonal anti-clathrin antibody followed by a secondary anti-goat IgG antibody coupled to fluorescein (green), CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary antibody anti-mouse IgG coupled to rhodamine (red), and Pet was detected using a rabbit anti-Pet polyclonal antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Confocal Microscopy

    Kidney cells transfected with ck8 are susceptible to cell damage caused by Pet. At 72 h posttransfection, MDCK cells transfected with ck8-gfp were treated with Pet (37 µg/ml) for 6 h. Untransfected and transfected cells were fixed, permeabilized, and stained with rhodamine-phalloidin for F actin (red). Pet was detected using a rabbit polyclonal anti-Pet antibody followed by anti-rabbit IgG secondary antibodies labeled with Cy5 (blue). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with fluorescein (green). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Kidney cells transfected with ck8 are susceptible to cell damage caused by Pet. At 72 h posttransfection, MDCK cells transfected with ck8-gfp were treated with Pet (37 µg/ml) for 6 h. Untransfected and transfected cells were fixed, permeabilized, and stained with rhodamine-phalloidin for F actin (red). Pet was detected using a rabbit polyclonal anti-Pet antibody followed by anti-rabbit IgG secondary antibodies labeled with Cy5 (blue). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with fluorescein (green). Slides were observed by confocal microscopy.

    Techniques Used: Transfection, Positron Emission Tomography, Staining, Labeling, Confocal Microscopy

    Pet and CK8 colocalize as a ligand-receptor complex in early endosomes of epithelial cells. HEp-2 cells were treated overnight with the CellLight early endosomes-RFP BacMam 2.0 reagent to mark Rab5b, a resident protein of early endosomes (red). Cells treated with Pet for 7, 8, 9, or 10 min were then washed, fixed, and permeabilized. CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary anti-mouse IgG antibody coupled to fluorescein (green), and Pet was detected using a rabbit polyclonal anti-Pet antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Pet and CK8 colocalize as a ligand-receptor complex in early endosomes of epithelial cells. HEp-2 cells were treated overnight with the CellLight early endosomes-RFP BacMam 2.0 reagent to mark Rab5b, a resident protein of early endosomes (red). Cells treated with Pet for 7, 8, 9, or 10 min were then washed, fixed, and permeabilized. CK8 was detected using a mouse monoclonal anti-CK8 antibody followed by a secondary anti-mouse IgG antibody coupled to fluorescein (green), and Pet was detected using a rabbit polyclonal anti-Pet antibody followed by a secondary anti-rabbit IgG antibody coupled to Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Confocal Microscopy

    Knockdown of CK8 prevents cell damage induced by Pet. (A) Knockdown of CK8 by siRNA. HEp-2 cells were silenced using siRNA for ck8 . Cells were lysed and analyzed by Western blotting using anti-CK8 monoclonal antibodies in comparison to normal cells. Results were analyzed by densitometry and plotted as the ratio of CK8 to actin. (B) Effect of Pet on CK8 knockdown cells. HEp-2 cells silenced using siRNA for ck8 were then treated with Pet (37 µg/ml) for 3 h, fixed, permeabilized, and stained with rhodaminated phalloidin (red). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with Cy5 (blue). Slides were observed by confocal microscopy.
    Figure Legend Snippet: Knockdown of CK8 prevents cell damage induced by Pet. (A) Knockdown of CK8 by siRNA. HEp-2 cells were silenced using siRNA for ck8 . Cells were lysed and analyzed by Western blotting using anti-CK8 monoclonal antibodies in comparison to normal cells. Results were analyzed by densitometry and plotted as the ratio of CK8 to actin. (B) Effect of Pet on CK8 knockdown cells. HEp-2 cells silenced using siRNA for ck8 were then treated with Pet (37 µg/ml) for 3 h, fixed, permeabilized, and stained with rhodaminated phalloidin (red). CK8 was detected using monoclonal anti-CK8 antibodies followed by a secondary anti-mouse IgG antibody labeled with Cy5 (blue). Slides were observed by confocal microscopy.

    Techniques Used: Positron Emission Tomography, Western Blot, Staining, Labeling, Confocal Microscopy

    24) Product Images from "Microcalcifications Drive Breast Cancer Occurrence and Development by Macrophage-Mediated Epithelial to Mesenchymal Transition"

    Article Title: Microcalcifications Drive Breast Cancer Occurrence and Development by Macrophage-Mediated Epithelial to Mesenchymal Transition

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20225633

    In Vitro model of BOLCs development. ( A ) Immunocytochemical analysis of MDA-MB-231 alone (MDA-MB-231/CTRL) shows no/rare vimentin-positive cells. ( B ) Several vimentin-positive MDA-MB-231 cells are present after co-culture with CO and activated monocytes. ( C ) Image displays starting point of MDA-MB-231 cells incubated with CO and activated monocytes. ( D ) After 10 days of culture, several large calcified nodules are present. ( A – D ) scale bars represent 100 µm ( E ) Western blot analysis for Tumor protein 53 (p53) and Pyruvate kinase Muscle (PKM) 2. ( F ) Western blot densitometric analysis (%) of p53 and PKM2 expression compared with the respective controls (MDA). ( G , H ) The ultrastructural aspect of MDA-MB-231 (CTRL). ( I – K ) Breast Osteoblast-like cells with hydroxyapatite intra-cytoplasmatic electron-dense granules (arrows) in MDA-MB-231 + Co + activated macrophages.
    Figure Legend Snippet: In Vitro model of BOLCs development. ( A ) Immunocytochemical analysis of MDA-MB-231 alone (MDA-MB-231/CTRL) shows no/rare vimentin-positive cells. ( B ) Several vimentin-positive MDA-MB-231 cells are present after co-culture with CO and activated monocytes. ( C ) Image displays starting point of MDA-MB-231 cells incubated with CO and activated monocytes. ( D ) After 10 days of culture, several large calcified nodules are present. ( A – D ) scale bars represent 100 µm ( E ) Western blot analysis for Tumor protein 53 (p53) and Pyruvate kinase Muscle (PKM) 2. ( F ) Western blot densitometric analysis (%) of p53 and PKM2 expression compared with the respective controls (MDA). ( G , H ) The ultrastructural aspect of MDA-MB-231 (CTRL). ( I – K ) Breast Osteoblast-like cells with hydroxyapatite intra-cytoplasmatic electron-dense granules (arrows) in MDA-MB-231 + Co + activated macrophages.

    Techniques Used: In Vitro, Multiple Displacement Amplification, Co-Culture Assay, Incubation, Western Blot, Expressing

    25) Product Images from "Enzyme-Linked Immunosorbent Assay for Detection of Filovirus Species-Specific Antibodies ▿"

    Article Title: Enzyme-Linked Immunosorbent Assay for Detection of Filovirus Species-Specific Antibodies ▿

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.00170-10

    IgG and IgM antibodies detected in human samples. OD values for specific IgG (a) and IgM (b) antibodies in the patient sera are shown. Sera from 21 individuals were analyzed at 1:1,000 dilutions. Naïve human sera (1:100 dilution) were used as a negative control. Each bar represents the average and standard deviation of data from independent experiments. Asterisks indicate statistically significant differences in OD values between the homologous antigen and all other antigens ( P
    Figure Legend Snippet: IgG and IgM antibodies detected in human samples. OD values for specific IgG (a) and IgM (b) antibodies in the patient sera are shown. Sera from 21 individuals were analyzed at 1:1,000 dilutions. Naïve human sera (1:100 dilution) were used as a negative control. Each bar represents the average and standard deviation of data from independent experiments. Asterisks indicate statistically significant differences in OD values between the homologous antigen and all other antigens ( P

    Techniques Used: Negative Control, Standard Deviation

    26) Product Images from "The potassium channel KCa3.1 constitutes a pharmacological target for neuroinflammation associated with ischemia/reperfusion stroke"

    Article Title: The potassium channel KCa3.1 constitutes a pharmacological target for neuroinflammation associated with ischemia/reperfusion stroke

    Journal: Journal of Cerebral Blood Flow & Metabolism

    doi: 10.1177/0271678X15611434

    K + -channel expression in acutely isolated microglia. (a) Kv1.3 current density increases in microglia from the infarct area after MCAO (28.8 ± 2.0 pA/pF, n = 19) and microglia isolated from the hippocampus following intracerebroventricular LPS injection (22.9 ± 16.6 pA/pF, n = 13) compared to microglia from wild-type control brains (5.0 ± 3.9 pA/pF, n = 16) or microglia from the contralateral side after MCAO (5.7 ± 4.4 pA/pF, n = 18). (b) Example current traces showing Kv1.3's characteristic use-dependence and sensitivity to the Kv1.3 blockers PAP-1 and ShK-L5. (c) Microglia from the contralateral (50.2 ± 35.4 pS/pF, n = 18) and ipsilateral side after MCAO (71.6 ± 34.9 pS/pF, n = 21), as well as microglia isolated from the hippocampus following intracerebroventricular LPS injection (84.0 ± 42.4 pS/pF, n = 13) show higher KCa3.1 current densities than microglia from wild-type control brains (29.7 ± 15.2 pS/pF, n = 16). (d) Example KCa3.1 current traces elicited by a ramp protocol showing the current's sensitive to 1 µM of the KCa3.1-selective blocker TRAM-34. (e) Microglia from both the contralateral side (7.8 ± 5.8 pA/pF, n = 18) and the infarct area (15.1 ± 10.2 pA/pF, n = 21) after MCAO show increased Kir current densities compared to those from wild-type (2.4 ± 2.4 pA/pF, n = 16) or LPS-injected brains (1.9 ± 2.9 pA/pF, n = 13). (f) Representative current traces showing a large Kir current, which was observable in some MCAO microglia, but not in microglia isolated from the hippocampus following intracerebroventricular LPS injection. Data are presented as mean ± S.D. Statistical significance was determined by Student's t -test.
    Figure Legend Snippet: K + -channel expression in acutely isolated microglia. (a) Kv1.3 current density increases in microglia from the infarct area after MCAO (28.8 ± 2.0 pA/pF, n = 19) and microglia isolated from the hippocampus following intracerebroventricular LPS injection (22.9 ± 16.6 pA/pF, n = 13) compared to microglia from wild-type control brains (5.0 ± 3.9 pA/pF, n = 16) or microglia from the contralateral side after MCAO (5.7 ± 4.4 pA/pF, n = 18). (b) Example current traces showing Kv1.3's characteristic use-dependence and sensitivity to the Kv1.3 blockers PAP-1 and ShK-L5. (c) Microglia from the contralateral (50.2 ± 35.4 pS/pF, n = 18) and ipsilateral side after MCAO (71.6 ± 34.9 pS/pF, n = 21), as well as microglia isolated from the hippocampus following intracerebroventricular LPS injection (84.0 ± 42.4 pS/pF, n = 13) show higher KCa3.1 current densities than microglia from wild-type control brains (29.7 ± 15.2 pS/pF, n = 16). (d) Example KCa3.1 current traces elicited by a ramp protocol showing the current's sensitive to 1 µM of the KCa3.1-selective blocker TRAM-34. (e) Microglia from both the contralateral side (7.8 ± 5.8 pA/pF, n = 18) and the infarct area (15.1 ± 10.2 pA/pF, n = 21) after MCAO show increased Kir current densities compared to those from wild-type (2.4 ± 2.4 pA/pF, n = 16) or LPS-injected brains (1.9 ± 2.9 pA/pF, n = 13). (f) Representative current traces showing a large Kir current, which was observable in some MCAO microglia, but not in microglia isolated from the hippocampus following intracerebroventricular LPS injection. Data are presented as mean ± S.D. Statistical significance was determined by Student's t -test.

    Techniques Used: Expressing, Isolation, Injection

    KCa3.1 and Kv1.3 are expressed on microglia/macrophages in human infarcts. (a) KCa3.1 staining in a 2–3-week-old infarct. KCa3.1 expression is localized to macrophages/microglia (M) and vascular endothelial (E) cells. (b) Fluorescent staining for a microglia/macrophage marker (MAC387) and KCa3.1. (c) Kv1.3 staining in a 14-day old-infarct. (d) Fluorescent staining for a microglia/macrophage marker (MAC387) and Kv1.3. All images are from 5-μm thick paraffin sections.
    Figure Legend Snippet: KCa3.1 and Kv1.3 are expressed on microglia/macrophages in human infarcts. (a) KCa3.1 staining in a 2–3-week-old infarct. KCa3.1 expression is localized to macrophages/microglia (M) and vascular endothelial (E) cells. (b) Fluorescent staining for a microglia/macrophage marker (MAC387) and KCa3.1. (c) Kv1.3 staining in a 14-day old-infarct. (d) Fluorescent staining for a microglia/macrophage marker (MAC387) and Kv1.3. All images are from 5-μm thick paraffin sections.

    Techniques Used: Staining, Expressing, Marker

    27) Product Images from "Tetraspanin CD82 interaction with cholesterol promotes extracellular vesicle–mediated release of ezrin to inhibit tumour cell movement"

    Article Title: Tetraspanin CD82 interaction with cholesterol promotes extracellular vesicle–mediated release of ezrin to inhibit tumour cell movement

    Journal: Journal of Extracellular Vesicles

    doi: 10.1080/20013078.2019.1692417

    CD82 and its cholesterol-binding differentially regulate cellular release of EVs. (a) Extracellular staining by filipin and Alexa488-conjugated CTxb in Du145 transfectants. Equal number of the cells were cultured on glass coverslips for 2 days, then fixed and labelled with filipin or Alexa488-conjugated CTxb. For filipin staining, intercellular regions were imaged. For CTxb staining, pericellular regions were imaged. Scale bar: 10 µm. (b) Distributions of Annexin V and Annexin A2 in Du145 transfectant cells. Alexa488-conjugated recombinant Annexin V was used for phosphatidylserine labelling, while Annexin-A2 Ab was used for Annexin-A2 staining. Scale bar: 10 μm. (c) Colocalization of Ezrin with GM1 or Annexin A2 in EVs. For Ezrin and GM1 co-staining, the cells were labelled with the Abs, Alexa488-conjugated CTxB and DAPI. For Ezrin and Annexin A2 co-staining, the cells were incubated sequentially with the primary Abs, Cy3-conjugated donkey anti-goat IgG, normal goat IgG and Alexa594-conjugated goat anti-mouse IgG. Images were obtained by confocal microscopy. Scale bar: 10 µm. (d) The cells were seeded in six-well plate at 50% confluence and cultured in DMEM containing 1% exosome-depleted FBS for 2 – 3 days. The culture supernatants were collected, spun at 2000 × g for 10 min to remove cell debris, and then analysed with NanoSight instrument for EV number and size. Data are presented as mean ± SD (n = 3 individual experiments). *: p
    Figure Legend Snippet: CD82 and its cholesterol-binding differentially regulate cellular release of EVs. (a) Extracellular staining by filipin and Alexa488-conjugated CTxb in Du145 transfectants. Equal number of the cells were cultured on glass coverslips for 2 days, then fixed and labelled with filipin or Alexa488-conjugated CTxb. For filipin staining, intercellular regions were imaged. For CTxb staining, pericellular regions were imaged. Scale bar: 10 µm. (b) Distributions of Annexin V and Annexin A2 in Du145 transfectant cells. Alexa488-conjugated recombinant Annexin V was used for phosphatidylserine labelling, while Annexin-A2 Ab was used for Annexin-A2 staining. Scale bar: 10 μm. (c) Colocalization of Ezrin with GM1 or Annexin A2 in EVs. For Ezrin and GM1 co-staining, the cells were labelled with the Abs, Alexa488-conjugated CTxB and DAPI. For Ezrin and Annexin A2 co-staining, the cells were incubated sequentially with the primary Abs, Cy3-conjugated donkey anti-goat IgG, normal goat IgG and Alexa594-conjugated goat anti-mouse IgG. Images were obtained by confocal microscopy. Scale bar: 10 µm. (d) The cells were seeded in six-well plate at 50% confluence and cultured in DMEM containing 1% exosome-depleted FBS for 2 – 3 days. The culture supernatants were collected, spun at 2000 × g for 10 min to remove cell debris, and then analysed with NanoSight instrument for EV number and size. Data are presented as mean ± SD (n = 3 individual experiments). *: p

    Techniques Used: Binding Assay, Staining, Cell Culture, Transfection, Recombinant, Incubation, Confocal Microscopy

    28) Product Images from "Structure function analysis of SH2D2A isoforms expressed in T cells reveals a crucial role for the proline rich region encoded by SH2D2A exon 7"

    Article Title: Structure function analysis of SH2D2A isoforms expressed in T cells reveals a crucial role for the proline rich region encoded by SH2D2A exon 7

    Journal: BMC Immunology

    doi: 10.1186/1471-2172-7-15

    Nuclear translocation of TSAd depends on aa239–334 . The SH2D2A-5 variant is excluded from the cell nucleus in Jurkat TAg cells: Confocal immunocytochemistry analysis of Jurkat TAg cells transiently transfected with pEF-HA-SH2D2A-2 and -5. The nuclear membrane protein LAP-2 was stained red (Cy™-3), whereas the HA-tagged recombinant proteins are stained green (Cy-2™). Cells were stimulated with anti-CD3 mAbs (OKT3) for 30 min. Horizontal sections are labelled Y, vertical sections are labelled Z (100× objective, 4× zoom).
    Figure Legend Snippet: Nuclear translocation of TSAd depends on aa239–334 . The SH2D2A-5 variant is excluded from the cell nucleus in Jurkat TAg cells: Confocal immunocytochemistry analysis of Jurkat TAg cells transiently transfected with pEF-HA-SH2D2A-2 and -5. The nuclear membrane protein LAP-2 was stained red (Cy™-3), whereas the HA-tagged recombinant proteins are stained green (Cy-2™). Cells were stimulated with anti-CD3 mAbs (OKT3) for 30 min. Horizontal sections are labelled Y, vertical sections are labelled Z (100× objective, 4× zoom).

    Techniques Used: Translocation Assay, Variant Assay, Immunocytochemistry, Transfection, Staining, Recombinant

    29) Product Images from "Murine homologue of the human KIAA1199 is implicated in hyaluronan binding and depolymerization"

    Article Title: Murine homologue of the human KIAA1199 is implicated in hyaluronan binding and depolymerization

    Journal: FEBS Open Bio

    doi: 10.1016/j.fob.2013.08.003

    Clathrin-specific HA depolymerization and cellular localization of mKiaa1199 in mKiaa1199/HEK293 cells. (A–C) Changes in HA depolymerization in mKiaa1199/HEK293 cells by knockdown of CHC (A), α-adaptin (B) or caveolin-1 (C) with siRNAs. As for controls, cells were transfected with control non-silencing siRNA. After incubation of the cells with [ 3 H]HA for 24 h, HA fragments in the media were analyzed by size exclusion chromatography. Efficiency of the knockdown was evaluated by immunoblotting (insets). Representative data from two siRNAs are shown. (D) Double immunostaining of mKiaa1199 (green) and CHC (red) with anti-KIAA1199 and anti-CHC antibodies in mKiaa1199/HEK293 cells. The inset shows a high-power view of the boxed area. Dotted lines show the plasma membrane. N, nucleus. Scale bar, 5 μm; scale bar for inset, 1 μm. (E) HA localization in mKiaa1199/HEK293 cells after incubation with biotin-labeled HA, followed by reaction with streptavidin conjugated to Alexa-Fluor 488. Arrows, HA-positive structures; N, nucleus. Scale bar, 5 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
    Figure Legend Snippet: Clathrin-specific HA depolymerization and cellular localization of mKiaa1199 in mKiaa1199/HEK293 cells. (A–C) Changes in HA depolymerization in mKiaa1199/HEK293 cells by knockdown of CHC (A), α-adaptin (B) or caveolin-1 (C) with siRNAs. As for controls, cells were transfected with control non-silencing siRNA. After incubation of the cells with [ 3 H]HA for 24 h, HA fragments in the media were analyzed by size exclusion chromatography. Efficiency of the knockdown was evaluated by immunoblotting (insets). Representative data from two siRNAs are shown. (D) Double immunostaining of mKiaa1199 (green) and CHC (red) with anti-KIAA1199 and anti-CHC antibodies in mKiaa1199/HEK293 cells. The inset shows a high-power view of the boxed area. Dotted lines show the plasma membrane. N, nucleus. Scale bar, 5 μm; scale bar for inset, 1 μm. (E) HA localization in mKiaa1199/HEK293 cells after incubation with biotin-labeled HA, followed by reaction with streptavidin conjugated to Alexa-Fluor 488. Arrows, HA-positive structures; N, nucleus. Scale bar, 5 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

    Techniques Used: Transfection, Incubation, Size-exclusion Chromatography, Double Immunostaining, Labeling

    HA-specific binding of recombinant mKiaa1199 protein expressed in mKiaa1199/HEK293 cells. (A) Binding assay of mKiaa1199 protein to different GAGs. Lysates of mKiaa1199/HEK293 cells were incubated with H 2 O (negative control) or unlabeled GAGs including HA (HA-H2), CSA, CSC, CSD, DS, Hep and HS. The samples were precipitated with CPC, and analyzed by immunoblotting with anti-KIAA1199 antibody. (B) Binding of mKiaa1199 to HA species with different molecular sizes. The lysates were incubated with H 2 O (negative control) or HA with different sizes including HA-H2, HA-M2, HA-L2, HA-S2 and HA-T2, and then subjected to immunoblotting with anti-KIAA1199 antibody.
    Figure Legend Snippet: HA-specific binding of recombinant mKiaa1199 protein expressed in mKiaa1199/HEK293 cells. (A) Binding assay of mKiaa1199 protein to different GAGs. Lysates of mKiaa1199/HEK293 cells were incubated with H 2 O (negative control) or unlabeled GAGs including HA (HA-H2), CSA, CSC, CSD, DS, Hep and HS. The samples were precipitated with CPC, and analyzed by immunoblotting with anti-KIAA1199 antibody. (B) Binding of mKiaa1199 to HA species with different molecular sizes. The lysates were incubated with H 2 O (negative control) or HA with different sizes including HA-H2, HA-M2, HA-L2, HA-S2 and HA-T2, and then subjected to immunoblotting with anti-KIAA1199 antibody.

    Techniques Used: Binding Assay, Recombinant, Incubation, Negative Control

    HA depolymerization by mKiaa1199 transfectants and determination of HA cleavage sites . (A) HEK293 cells were transiently transfected with empty vector (Mock) or vector containing hKIAA1199 or mKiaa1199 cDNA. The expression of hKIAA1199 and mKiaa1199 proteins in each transfectant was assessed by immunoblotting using anti-KIAA1199 monoclonal antibody (inset). Mock and mKiaa1199 transfectants were incubated with [ 3 H]HA for 48 h, and HA depolymerization was then examined by size exclusion chromatography. GAPDH, a loading control. (B) Determination of the non-reducing terminal sugar of depolymerized HA. [ 3 H]HA fragments obtained from the culture media were incubated with β- N -acetylglucosaminidase (open circles) or β-glucuronidase followed by incubation with β- N -acetylglucosaminidase (closed circles), and then applied to Sephadex G-25 column chromatogram.
    Figure Legend Snippet: HA depolymerization by mKiaa1199 transfectants and determination of HA cleavage sites . (A) HEK293 cells were transiently transfected with empty vector (Mock) or vector containing hKIAA1199 or mKiaa1199 cDNA. The expression of hKIAA1199 and mKiaa1199 proteins in each transfectant was assessed by immunoblotting using anti-KIAA1199 monoclonal antibody (inset). Mock and mKiaa1199 transfectants were incubated with [ 3 H]HA for 48 h, and HA depolymerization was then examined by size exclusion chromatography. GAPDH, a loading control. (B) Determination of the non-reducing terminal sugar of depolymerized HA. [ 3 H]HA fragments obtained from the culture media were incubated with β- N -acetylglucosaminidase (open circles) or β-glucuronidase followed by incubation with β- N -acetylglucosaminidase (closed circles), and then applied to Sephadex G-25 column chromatogram.

    Techniques Used: Transfection, Plasmid Preparation, Expressing, Incubation, Size-exclusion Chromatography

    30) Product Images from "SCCRO3 (DCUN1D3) Antagonizes the Neddylation and Oncogenic Activity of SCCRO (DCUN1D1) *"

    Article Title: SCCRO3 (DCUN1D3) Antagonizes the Neddylation and Oncogenic Activity of SCCRO (DCUN1D1) *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.585505

    SCCRO3 competes with SCCRO for subcellular localization of cullins. A , representative images from fluorescence microscopy ( scale bar represents 20 μm) showing U2OS cells co-transfected with Myc- Cul1 , the indicated HA-tagged constructs and varying concentrations of untagged SCCRO ( fifth and sixth rows ) stained with FITC-conjugated anti-HA ( first column ) and Cy3-conjugated anti-Myc antibody ( second column ) or DAPI ( third column ). Merged images are shown ( fourth column ). Myc-Cul1 co-localized with HA-SCCRO ( first row ) in the nucleus and HA-SCCRO3 at the membrane ( second row ). HA-SCCRO3 G2A loses membrane localization, and Myc-Cul1 is primarily nuclear in these cells ( third row ). HA-SCCRO3 DAD retains membrane localization, but HA-Cul1 does not co-localize with it ( fourth row ). Co-transfection of increasing concentrations of SCCRO with HA-SCCRO3 results in progressive translocation of Myc-Cul1 from the membrane to the nucleus ( fifth and sixth rows ). B , graph showing subcellular distribution of HA-Cul1 from experiment above at varying concentrations of SCRRO and SCCRO3. C , representative fluorescence microscopic images of U2OS cells transfected with scrambled shRNA, or shRNA against SCCRO or SCCRO3 stained for Cul1 using antibody against Cul1 ( first column ) and DAPI ( second column ) and merged ( third column ). Knockdown of SCCRO resulted in localization of a fraction of Cul1 to the cell membrane ( second row ). Knockdown of SCCRO3 increased the proportion of Cul1 in the nucleus ( third row ). D , Western blot analysis of lysates from U2OS cells expressing shRNA against SCCRO (shRNA1 and shRNA2) showing reduced levels of SCCRO ( third row ), but not SCCRO3 ( fourth row ), and a decrease in the proportion of neddylated Cul1 and Cul3. E , Western blot analysis of lysates from U2OS cells expressing shRNA against SCCRO3 (shRNA1 and shRNA2) showing decreased SCCRO3, but not SCCRO expression, and an increase in the proportion of neddylated Cul1 and Cul3. The numbers below the blots are the ratios of neddylated to nonneddylated cullins.
    Figure Legend Snippet: SCCRO3 competes with SCCRO for subcellular localization of cullins. A , representative images from fluorescence microscopy ( scale bar represents 20 μm) showing U2OS cells co-transfected with Myc- Cul1 , the indicated HA-tagged constructs and varying concentrations of untagged SCCRO ( fifth and sixth rows ) stained with FITC-conjugated anti-HA ( first column ) and Cy3-conjugated anti-Myc antibody ( second column ) or DAPI ( third column ). Merged images are shown ( fourth column ). Myc-Cul1 co-localized with HA-SCCRO ( first row ) in the nucleus and HA-SCCRO3 at the membrane ( second row ). HA-SCCRO3 G2A loses membrane localization, and Myc-Cul1 is primarily nuclear in these cells ( third row ). HA-SCCRO3 DAD retains membrane localization, but HA-Cul1 does not co-localize with it ( fourth row ). Co-transfection of increasing concentrations of SCCRO with HA-SCCRO3 results in progressive translocation of Myc-Cul1 from the membrane to the nucleus ( fifth and sixth rows ). B , graph showing subcellular distribution of HA-Cul1 from experiment above at varying concentrations of SCRRO and SCCRO3. C , representative fluorescence microscopic images of U2OS cells transfected with scrambled shRNA, or shRNA against SCCRO or SCCRO3 stained for Cul1 using antibody against Cul1 ( first column ) and DAPI ( second column ) and merged ( third column ). Knockdown of SCCRO resulted in localization of a fraction of Cul1 to the cell membrane ( second row ). Knockdown of SCCRO3 increased the proportion of Cul1 in the nucleus ( third row ). D , Western blot analysis of lysates from U2OS cells expressing shRNA against SCCRO (shRNA1 and shRNA2) showing reduced levels of SCCRO ( third row ), but not SCCRO3 ( fourth row ), and a decrease in the proportion of neddylated Cul1 and Cul3. E , Western blot analysis of lysates from U2OS cells expressing shRNA against SCCRO3 (shRNA1 and shRNA2) showing decreased SCCRO3, but not SCCRO expression, and an increase in the proportion of neddylated Cul1 and Cul3. The numbers below the blots are the ratios of neddylated to nonneddylated cullins.

    Techniques Used: Fluorescence, Microscopy, Transfection, Construct, Staining, Cotransfection, Translocation Assay, shRNA, Western Blot, Expressing

    31) Product Images from "The Ca2+-Activated K+ Channel KCa3.1 as a Potential New Target for the Prevention of Allograft Vasculopathy"

    Article Title: The Ca2+-Activated K+ Channel KCa3.1 as a Potential New Target for the Prevention of Allograft Vasculopathy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0081006

    KCa3.1 staining localizes to smooth muscle cells, T cells and macrophages. Double fluorescent immunostaining for KCa3.1 and α-SMA, T cells (CD43) and macrophages (ED1, CD68) in orthotopic rat aorta transplants harvested 80 or 120 days after transplantation. The boxed areas in the H E stained vessels on the right show the location where the fluorescent images were taken.
    Figure Legend Snippet: KCa3.1 staining localizes to smooth muscle cells, T cells and macrophages. Double fluorescent immunostaining for KCa3.1 and α-SMA, T cells (CD43) and macrophages (ED1, CD68) in orthotopic rat aorta transplants harvested 80 or 120 days after transplantation. The boxed areas in the H E stained vessels on the right show the location where the fluorescent images were taken.

    Techniques Used: Staining, Immunostaining, Transplantation Assay

    32) Product Images from "Role of Phosphatidylinositol 4,5-Bisphosphate in Regulating EHD2 Plasma Membrane Localization"

    Article Title: Role of Phosphatidylinositol 4,5-Bisphosphate in Regulating EHD2 Plasma Membrane Localization

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0074519

    Acute PLD inhibition reduces EHD2 protein levels, but does not dissociate EHD2 from membranes. (A) HeLa cells were treated with DMSO (control), or 15 µM each CAY10593 and CAY10594 for 4 h, or 25 µM each CAY10593 and CAY10594 for 45 min. The cells were then lysed, and equal amounts of proteins were separated by SDS-PAGE and subjected to immunoblotting with antibodies against EHD2, caveolin1 or actin. The immunoblots are representative of seven independent experiments. (B) HeLa cells were pre-treated with 100 µM leupeptin or with 10 µM lactacystin for 3 h. The cells were then treated with fresh 100 µM leupeptin or 10 µM lactacystin along with either DMSO, 15 µM each CAY10593 and CAY10594 (for 4 h), or 25 µM each CAY10593 and CAY10594 (for 45 min). Following treatment, the cells were lysed, and equal amounts of proteins were separated by SDS-PAGE and subjected to immunoblotting with antibodies against EHD2, caveolin1 or actin. The immunoblots are representative of three experiments conducted with leupeptin, and two experiments done with lactacystin. (C) HeLa cells were treated as in (A). The cells were then homogenized and post-nuclear supernatants were separated into cytosol and membrane fractions by ultracentrifugation. The samples were resolved by SDS-PAGE and immunoblotted with antibodies against EHD2, caveolin1 or cytochrome C (cytosolic fraction control). The immunoblots are representative of two independent experiments.
    Figure Legend Snippet: Acute PLD inhibition reduces EHD2 protein levels, but does not dissociate EHD2 from membranes. (A) HeLa cells were treated with DMSO (control), or 15 µM each CAY10593 and CAY10594 for 4 h, or 25 µM each CAY10593 and CAY10594 for 45 min. The cells were then lysed, and equal amounts of proteins were separated by SDS-PAGE and subjected to immunoblotting with antibodies against EHD2, caveolin1 or actin. The immunoblots are representative of seven independent experiments. (B) HeLa cells were pre-treated with 100 µM leupeptin or with 10 µM lactacystin for 3 h. The cells were then treated with fresh 100 µM leupeptin or 10 µM lactacystin along with either DMSO, 15 µM each CAY10593 and CAY10594 (for 4 h), or 25 µM each CAY10593 and CAY10594 (for 45 min). Following treatment, the cells were lysed, and equal amounts of proteins were separated by SDS-PAGE and subjected to immunoblotting with antibodies against EHD2, caveolin1 or actin. The immunoblots are representative of three experiments conducted with leupeptin, and two experiments done with lactacystin. (C) HeLa cells were treated as in (A). The cells were then homogenized and post-nuclear supernatants were separated into cytosol and membrane fractions by ultracentrifugation. The samples were resolved by SDS-PAGE and immunoblotted with antibodies against EHD2, caveolin1 or cytochrome C (cytosolic fraction control). The immunoblots are representative of two independent experiments.

    Techniques Used: Inhibition, SDS Page, Western Blot

    EHD2 localizes to the plasma membrane independent of its interaction partners, syndapin2 and EHBP1. HeLa cells were treated with non-targeting control-, syndapin2- or EHBP1-siRNA for 72 h. Cells were then lysed, and equal amounts of proteins were separated by SDS-PAGE and subjected to immunoblotting with antibodies against syndapin2, EHBP1, EHD2, caveolin1 or actin (A). For confocal microscopy, cells growing on coverslips were fixed and stained with antibodies against EHD2 (B–D) or caveolin1 (E–G). Bar, 10 µm. The immunoblots and confocal micrographs are representative of three independent experiments.
    Figure Legend Snippet: EHD2 localizes to the plasma membrane independent of its interaction partners, syndapin2 and EHBP1. HeLa cells were treated with non-targeting control-, syndapin2- or EHBP1-siRNA for 72 h. Cells were then lysed, and equal amounts of proteins were separated by SDS-PAGE and subjected to immunoblotting with antibodies against syndapin2, EHBP1, EHD2, caveolin1 or actin (A). For confocal microscopy, cells growing on coverslips were fixed and stained with antibodies against EHD2 (B–D) or caveolin1 (E–G). Bar, 10 µm. The immunoblots and confocal micrographs are representative of three independent experiments.

    Techniques Used: SDS Page, Confocal Microscopy, Staining, Western Blot

    33) Product Images from "Structure function analysis of SH2D2A isoforms expressed in T cells reveals a crucial role for the proline rich region encoded by SH2D2A exon 7"

    Article Title: Structure function analysis of SH2D2A isoforms expressed in T cells reveals a crucial role for the proline rich region encoded by SH2D2A exon 7

    Journal: BMC Immunology

    doi: 10.1186/1471-2172-7-15

    Nuclear translocation of TSAd depends on aa239–334 . The SH2D2A-5 variant is excluded from the cell nucleus in Jurkat TAg cells: Confocal immunocytochemistry analysis of Jurkat TAg cells transiently transfected with pEF-HA-SH2D2A-2 and -5. The nuclear membrane protein LAP-2 was stained red (Cy™-3), whereas the HA-tagged recombinant proteins are stained green (Cy-2™). Cells were stimulated with anti-CD3 mAbs (OKT3) for 30 min. Horizontal sections are labelled Y, vertical sections are labelled Z (100× objective, 4× zoom).
    Figure Legend Snippet: Nuclear translocation of TSAd depends on aa239–334 . The SH2D2A-5 variant is excluded from the cell nucleus in Jurkat TAg cells: Confocal immunocytochemistry analysis of Jurkat TAg cells transiently transfected with pEF-HA-SH2D2A-2 and -5. The nuclear membrane protein LAP-2 was stained red (Cy™-3), whereas the HA-tagged recombinant proteins are stained green (Cy-2™). Cells were stimulated with anti-CD3 mAbs (OKT3) for 30 min. Horizontal sections are labelled Y, vertical sections are labelled Z (100× objective, 4× zoom).

    Techniques Used: Translocation Assay, Variant Assay, Immunocytochemistry, Transfection, Staining, Recombinant

    34) Product Images from "Factors Determining the Risk of Inadvertent Retroviral Transduction of Male Germ Cells After In Utero Gene Transfer in Sheep"

    Article Title: Factors Determining the Risk of Inadvertent Retroviral Transduction of Male Germ Cells After In Utero Gene Transfer in Sheep

    Journal: Human Gene Therapy

    doi: 10.1089/hum.2007.120

    GCs migrating within the rete testes and interstitial space express SSEA-1
    Figure Legend Snippet: GCs migrating within the rete testes and interstitial space express SSEA-1

    Techniques Used:

    Migrating GCs in the fetal testes express SSEA-1. ( A ) High-power (× 60 objective) magnification showing high levels of SSEA-1 expression in migrating GCs (M), weak SSEA-1 expression in partially compartmentalized GCs (P), and undetectable
    Figure Legend Snippet: Migrating GCs in the fetal testes express SSEA-1. ( A ) High-power (× 60 objective) magnification showing high levels of SSEA-1 expression in migrating GCs (M), weak SSEA-1 expression in partially compartmentalized GCs (P), and undetectable

    Techniques Used: Expressing

    35) Product Images from "Diverse Requirements for Src-Family Tyrosine Kinases Distinguish Chlamydial Species"

    Article Title: Diverse Requirements for Src-Family Tyrosine Kinases Distinguish Chlamydial Species

    Journal: mBio

    doi: 10.1128/mBio.00031-11

    Trafficking of nascent chlamydial inclusions to the MTOC is species specific. (A) L929 cells were infected with C. trachomatis L2, C. caviae GPIC, C. muridarum MoPn, or C. pneumoniae AR39. Five hours postinfection, the cells were fixed and labeled with anti-EB antisera (green) and anti-γ-tubulin antibodies (red) followed by DyLight 488 and 594 secondary antibodies as well as DRAQ5 DNA stain (blue). Arrowheads indicate early inclusions converged at the MTOC. Bar, 10 µm. (B) The percentage of infected cells with EBs clustered at the MTOC was determined using confocal fluorescent microscopy. C. trachomatis L2 and C. pneumoniae consistently traffic to the centrosome, whereas C. caviae and C. muridarum do not. One hundred cells were counted per sample ( n = 3); error bars show standard deviations. Cpn, C. pneumoniae .
    Figure Legend Snippet: Trafficking of nascent chlamydial inclusions to the MTOC is species specific. (A) L929 cells were infected with C. trachomatis L2, C. caviae GPIC, C. muridarum MoPn, or C. pneumoniae AR39. Five hours postinfection, the cells were fixed and labeled with anti-EB antisera (green) and anti-γ-tubulin antibodies (red) followed by DyLight 488 and 594 secondary antibodies as well as DRAQ5 DNA stain (blue). Arrowheads indicate early inclusions converged at the MTOC. Bar, 10 µm. (B) The percentage of infected cells with EBs clustered at the MTOC was determined using confocal fluorescent microscopy. C. trachomatis L2 and C. pneumoniae consistently traffic to the centrosome, whereas C. caviae and C. muridarum do not. One hundred cells were counted per sample ( n = 3); error bars show standard deviations. Cpn, C. pneumoniae .

    Techniques Used: Infection, Labeling, Staining, Microscopy

    36) Product Images from "The AP-2 Adaptor ?2 Appendage Scaffolds Alternate Cargo Endocytosis"

    Article Title: The AP-2 Adaptor ?2 Appendage Scaffolds Alternate Cargo Endocytosis

    Journal: Molecular Biology of the Cell

    doi: 10.1091/mbc.E08-07-0712

    AP-2 knocked down cell ultrastructure. HeLa SS6 cells either mock transfected (A) or transfected with α subunit siRNA oligonucleotides (B–D) were briefly sonicated to prepare cell cortices, fixed, labeled with anti-transferrin receptor mAb H68.4, and then 15-nm gold-conjugated anti-mouse secondary antibodies to visualize extra-lattice receptors. Arrows indicate gold-labeled receptors.
    Figure Legend Snippet: AP-2 knocked down cell ultrastructure. HeLa SS6 cells either mock transfected (A) or transfected with α subunit siRNA oligonucleotides (B–D) were briefly sonicated to prepare cell cortices, fixed, labeled with anti-transferrin receptor mAb H68.4, and then 15-nm gold-conjugated anti-mouse secondary antibodies to visualize extra-lattice receptors. Arrows indicate gold-labeled receptors.

    Techniques Used: Transfection, Sonication, Labeling

    37) Product Images from "Dasatinib modulates sensitivity to pemetrexed in malignant pleural mesothelioma cell lines"

    Article Title: Dasatinib modulates sensitivity to pemetrexed in malignant pleural mesothelioma cell lines

    Journal: Oncotarget

    doi: 10.18632/oncotarget.10428

    Basal TS, pSRC and SRC localization and expression in MPM cells In all MPM cell lines analyzed, the subcellular distribution of both TS and pSRC was detected by confocal microscopy after immunostaining with anti-TS (FITC-green, left panel) and anti-pSRC (FITC-green, right panel). Propidium iodide was used to counter stain nuclei, and the images were overlaid to determine both TS and pSRC localization within the cell. Confocal series images were taken on an inverted Zeiss LSM510 microscope equipped with a Plan-Apochromat 63X/1.4 oil immersion Ph 3 objective (Oberkochen, Germany). Scale bars: 10 μm. ( A ). Basal TS and SRC genes expression was determined by means of quantitative Real Time PCR ( B ) and TS, SRC and pSRC protein expression was also detected using immunoblotting ( C ). BACT served as housekeeping control.
    Figure Legend Snippet: Basal TS, pSRC and SRC localization and expression in MPM cells In all MPM cell lines analyzed, the subcellular distribution of both TS and pSRC was detected by confocal microscopy after immunostaining with anti-TS (FITC-green, left panel) and anti-pSRC (FITC-green, right panel). Propidium iodide was used to counter stain nuclei, and the images were overlaid to determine both TS and pSRC localization within the cell. Confocal series images were taken on an inverted Zeiss LSM510 microscope equipped with a Plan-Apochromat 63X/1.4 oil immersion Ph 3 objective (Oberkochen, Germany). Scale bars: 10 μm. ( A ). Basal TS and SRC genes expression was determined by means of quantitative Real Time PCR ( B ) and TS, SRC and pSRC protein expression was also detected using immunoblotting ( C ). BACT served as housekeeping control.

    Techniques Used: Expressing, Confocal Microscopy, Immunostaining, Staining, Microscopy, Real-time Polymerase Chain Reaction

    38) Product Images from "CIN85 Modulates the Down-regulation of Fc?RIIa Expression and Function by c-Cbl in a PKC-dependent Manner in Human Neutrophils *"

    Article Title: CIN85 Modulates the Down-regulation of Fc?RIIa Expression and Function by c-Cbl in a PKC-dependent Manner in Human Neutrophils *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.213660

    CIN85 is involved in the late steps of FcγRIIa internalization. Bt 2 cAMP-differentiated PLB-985 cells were transfected with a negative control siRNA or an siRNA against CIN85, as described under “Experimental Procedures. A , the membrane expression of FcγRIIa was monitored by flow cytometry. Transfected cells were stimulated for the indicated times by cross-linking FcγRIIa with a goat anti-mouse F(ab′) 2 anti- F(ab′) 2 (25 μg/ml) at 37 °C. The stimulations were terminated by transferring the tubes to an ice bath, followed by centrifugation at 400 × g for 2 min at 4 °C. The cell pellets were washed in cold HBSS containing 0.005% BSA and incubated with FITC-labeled goat anti-mouse Fcγ specific IgG (diluted 1:100 in HBSS/BSA) for 30 min on ice. Cells were then washed twice in HBSS/BSA and analyzed by flow cytometry. IgG2b was used as an isotype-matched negative control. The data shown are the quantification of the mean fluorescence intensity of three independent experiments where FcγRIIa levels were normalized to samples without IV.3 cross-linking (100% expression level). B , transfected PLB-985 (10 7 /ml) were incubated with phycoerythrin-conjugated IV.3 (1 μg/ml) for 10 min, washed and plated as described under “Experimental Procedures.” Anti-mouse F(ab′) 2 was added in the environment chamber, and cells were visualized live by confocal microscopy for the indicated times. For each condition, at least 25 cells were analyzed. These pictures are representative of three independent cross-linking experiments. Error bars , S.E.
    Figure Legend Snippet: CIN85 is involved in the late steps of FcγRIIa internalization. Bt 2 cAMP-differentiated PLB-985 cells were transfected with a negative control siRNA or an siRNA against CIN85, as described under “Experimental Procedures. A , the membrane expression of FcγRIIa was monitored by flow cytometry. Transfected cells were stimulated for the indicated times by cross-linking FcγRIIa with a goat anti-mouse F(ab′) 2 anti- F(ab′) 2 (25 μg/ml) at 37 °C. The stimulations were terminated by transferring the tubes to an ice bath, followed by centrifugation at 400 × g for 2 min at 4 °C. The cell pellets were washed in cold HBSS containing 0.005% BSA and incubated with FITC-labeled goat anti-mouse Fcγ specific IgG (diluted 1:100 in HBSS/BSA) for 30 min on ice. Cells were then washed twice in HBSS/BSA and analyzed by flow cytometry. IgG2b was used as an isotype-matched negative control. The data shown are the quantification of the mean fluorescence intensity of three independent experiments where FcγRIIa levels were normalized to samples without IV.3 cross-linking (100% expression level). B , transfected PLB-985 (10 7 /ml) were incubated with phycoerythrin-conjugated IV.3 (1 μg/ml) for 10 min, washed and plated as described under “Experimental Procedures.” Anti-mouse F(ab′) 2 was added in the environment chamber, and cells were visualized live by confocal microscopy for the indicated times. For each condition, at least 25 cells were analyzed. These pictures are representative of three independent cross-linking experiments. Error bars , S.E.

    Techniques Used: Transfection, Negative Control, Expressing, Flow Cytometry, Cytometry, Transferring, Centrifugation, Incubation, Labeling, Fluorescence, Confocal Microscopy

    39) Product Images from "The potassium channel KCa3.1 constitutes a pharmacological target for neuroinflammation associated with ischemia/reperfusion stroke"

    Article Title: The potassium channel KCa3.1 constitutes a pharmacological target for neuroinflammation associated with ischemia/reperfusion stroke

    Journal: Journal of Cerebral Blood Flow & Metabolism

    doi: 10.1177/0271678X15611434

    KCa3.1 expression on microglia/macrophages in mouse infarcts. (a) Fluorescent staining for KCa3.1 and CD68 in the infarct area of KCa3.1 −/− (a) and wild-type (b) mice five days after MCAO with reperfusion. (c) Higher magnification image showing that KCa3.1 and CD68 do not colocalize. All panels are confocal images from 5-μm thick paraffin sections.
    Figure Legend Snippet: KCa3.1 expression on microglia/macrophages in mouse infarcts. (a) Fluorescent staining for KCa3.1 and CD68 in the infarct area of KCa3.1 −/− (a) and wild-type (b) mice five days after MCAO with reperfusion. (c) Higher magnification image showing that KCa3.1 and CD68 do not colocalize. All panels are confocal images from 5-μm thick paraffin sections.

    Techniques Used: Expressing, Staining, Mouse Assay

    Genetic KCa3.1 deletion and pharmacological blockade with TRAM-34 reduce microglia activation and cytokine production. (a) Microglia/macrophage activation/proliferation/infiltration as determined by the ratio of the Iba-1 positive cells in the ipsi- versus contralateral hemisphere from the 4- and 6-mm slices from all animals in vehicle-treated wild-type mice (n = 11), vehicle-treated KCa3.1 −/− mice (n = 17) and wild-type mice treated with TRAM-34 at 40 mg/kg twice daily starting 12 h after reperfusion (n = 11). (b) Brain cytokine concentrations in the ipsi- and contralateral side eight days after MCAO from vehicle-treated wild-type mice, KCa3.1 −/− mice, TRAM-34-treated wild-type mice and vehicle-treated sham-operated wild-type mice (n = 3 in every case). * p
    Figure Legend Snippet: Genetic KCa3.1 deletion and pharmacological blockade with TRAM-34 reduce microglia activation and cytokine production. (a) Microglia/macrophage activation/proliferation/infiltration as determined by the ratio of the Iba-1 positive cells in the ipsi- versus contralateral hemisphere from the 4- and 6-mm slices from all animals in vehicle-treated wild-type mice (n = 11), vehicle-treated KCa3.1 −/− mice (n = 17) and wild-type mice treated with TRAM-34 at 40 mg/kg twice daily starting 12 h after reperfusion (n = 11). (b) Brain cytokine concentrations in the ipsi- and contralateral side eight days after MCAO from vehicle-treated wild-type mice, KCa3.1 −/− mice, TRAM-34-treated wild-type mice and vehicle-treated sham-operated wild-type mice (n = 3 in every case). * p

    Techniques Used: Activation Assay, Mouse Assay

    K + -channel expression in acutely isolated microglia. (a) Kv1.3 current density increases in microglia from the infarct area after MCAO (28.8 ± 2.0 pA/pF, n = 19) and microglia isolated from the hippocampus following intracerebroventricular LPS injection (22.9 ± 16.6 pA/pF, n = 13) compared to microglia from wild-type control brains (5.0 ± 3.9 pA/pF, n = 16) or microglia from the contralateral side after MCAO (5.7 ± 4.4 pA/pF, n = 18). (b) Example current traces showing Kv1.3's characteristic use-dependence and sensitivity to the Kv1.3 blockers PAP-1 and ShK-L5. (c) Microglia from the contralateral (50.2 ± 35.4 pS/pF, n = 18) and ipsilateral side after MCAO (71.6 ± 34.9 pS/pF, n = 21), as well as microglia isolated from the hippocampus following intracerebroventricular LPS injection (84.0 ± 42.4 pS/pF, n = 13) show higher KCa3.1 current densities than microglia from wild-type control brains (29.7 ± 15.2 pS/pF, n = 16). (d) Example KCa3.1 current traces elicited by a ramp protocol showing the current's sensitive to 1 µM of the KCa3.1-selective blocker TRAM-34. (e) Microglia from both the contralateral side (7.8 ± 5.8 pA/pF, n = 18) and the infarct area (15.1 ± 10.2 pA/pF, n = 21) after MCAO show increased Kir current densities compared to those from wild-type (2.4 ± 2.4 pA/pF, n = 16) or LPS-injected brains (1.9 ± 2.9 pA/pF, n = 13). (f) Representative current traces showing a large Kir current, which was observable in some MCAO microglia, but not in microglia isolated from the hippocampus following intracerebroventricular LPS injection. Data are presented as mean ± S.D. Statistical significance was determined by Student's t -test.
    Figure Legend Snippet: K + -channel expression in acutely isolated microglia. (a) Kv1.3 current density increases in microglia from the infarct area after MCAO (28.8 ± 2.0 pA/pF, n = 19) and microglia isolated from the hippocampus following intracerebroventricular LPS injection (22.9 ± 16.6 pA/pF, n = 13) compared to microglia from wild-type control brains (5.0 ± 3.9 pA/pF, n = 16) or microglia from the contralateral side after MCAO (5.7 ± 4.4 pA/pF, n = 18). (b) Example current traces showing Kv1.3's characteristic use-dependence and sensitivity to the Kv1.3 blockers PAP-1 and ShK-L5. (c) Microglia from the contralateral (50.2 ± 35.4 pS/pF, n = 18) and ipsilateral side after MCAO (71.6 ± 34.9 pS/pF, n = 21), as well as microglia isolated from the hippocampus following intracerebroventricular LPS injection (84.0 ± 42.4 pS/pF, n = 13) show higher KCa3.1 current densities than microglia from wild-type control brains (29.7 ± 15.2 pS/pF, n = 16). (d) Example KCa3.1 current traces elicited by a ramp protocol showing the current's sensitive to 1 µM of the KCa3.1-selective blocker TRAM-34. (e) Microglia from both the contralateral side (7.8 ± 5.8 pA/pF, n = 18) and the infarct area (15.1 ± 10.2 pA/pF, n = 21) after MCAO show increased Kir current densities compared to those from wild-type (2.4 ± 2.4 pA/pF, n = 16) or LPS-injected brains (1.9 ± 2.9 pA/pF, n = 13). (f) Representative current traces showing a large Kir current, which was observable in some MCAO microglia, but not in microglia isolated from the hippocampus following intracerebroventricular LPS injection. Data are presented as mean ± S.D. Statistical significance was determined by Student's t -test.

    Techniques Used: Expressing, Isolation, Injection

    KCa3.1 and Kv1.3 are expressed on microglia/macrophages in human infarcts. (a) KCa3.1 staining in a 2–3-week-old infarct. KCa3.1 expression is localized to macrophages/microglia (M) and vascular endothelial (E) cells. (b) Fluorescent staining for a microglia/macrophage marker (MAC387) and KCa3.1. (c) Kv1.3 staining in a 14-day old-infarct. (d) Fluorescent staining for a microglia/macrophage marker (MAC387) and Kv1.3. All images are from 5-μm thick paraffin sections.
    Figure Legend Snippet: KCa3.1 and Kv1.3 are expressed on microglia/macrophages in human infarcts. (a) KCa3.1 staining in a 2–3-week-old infarct. KCa3.1 expression is localized to macrophages/microglia (M) and vascular endothelial (E) cells. (b) Fluorescent staining for a microglia/macrophage marker (MAC387) and KCa3.1. (c) Kv1.3 staining in a 14-day old-infarct. (d) Fluorescent staining for a microglia/macrophage marker (MAC387) and Kv1.3. All images are from 5-μm thick paraffin sections.

    Techniques Used: Staining, Expressing, Marker

    Genetic KCa3.1 deletion and pharmacological blockade with TRAM-34 reduce infarction and improve neurological deficit. (a) NeuN-defined infarct area in mice subjected to 60 min of MCAO followed by eight days of reperfusion in brain slices 2, 4, 6, and 8 mm from the frontal pole. Shown are vehicle-treated wild-type mice (n = 11), vehicle-treated KCa3.1 −/− mice (n = 17) and wild-type mice treated with TRAM-34 at 40 mg/kg twice daily starting 12 h after reperfusion (n = 11). * p
    Figure Legend Snippet: Genetic KCa3.1 deletion and pharmacological blockade with TRAM-34 reduce infarction and improve neurological deficit. (a) NeuN-defined infarct area in mice subjected to 60 min of MCAO followed by eight days of reperfusion in brain slices 2, 4, 6, and 8 mm from the frontal pole. Shown are vehicle-treated wild-type mice (n = 11), vehicle-treated KCa3.1 −/− mice (n = 17) and wild-type mice treated with TRAM-34 at 40 mg/kg twice daily starting 12 h after reperfusion (n = 11). * p

    Techniques Used: Mouse Assay

    40) Product Images from "UBE2I (UBC9), a SUMO-Conjugating Enzyme, Localizes to Nuclear Speckles and Stimulates Transcription in Mouse Oocytes 1"

    Article Title: UBE2I (UBC9), a SUMO-Conjugating Enzyme, Localizes to Nuclear Speckles and Stimulates Transcription in Mouse Oocytes 1

    Journal:

    doi: 10.1095/biolreprod.108.070474

    Subcellular localization of UBE2I, SUMO1, and SUMO2 and SUMO3 in the nucleus of oocytes at different stages of development. A ) Day 13 meiotically incompetent oocyte. B ) Fully grown NSN oocyte. C ) Fully grown SN oocyte. In each panel, oocytes were stained
    Figure Legend Snippet: Subcellular localization of UBE2I, SUMO1, and SUMO2 and SUMO3 in the nucleus of oocytes at different stages of development. A ) Day 13 meiotically incompetent oocyte. B ) Fully grown NSN oocyte. C ) Fully grown SN oocyte. In each panel, oocytes were stained

    Techniques Used: Staining

    Subcellular localization of UBE2I in oocytes and preimplantation embryos. A ) UBE2I was stained with the anti-UBE2I antibody (red). The nuclei were stained with SYTOX green (green). a – h ) Day 13 incompetent oocyte ( a ), GV oocyte ( b ), GVBD oocyte
    Figure Legend Snippet: Subcellular localization of UBE2I in oocytes and preimplantation embryos. A ) UBE2I was stained with the anti-UBE2I antibody (red). The nuclei were stained with SYTOX green (green). a – h ) Day 13 incompetent oocyte ( a ), GV oocyte ( b ), GVBD oocyte

    Techniques Used: Staining

    UBE2I stimulates BrUTP incorporation independent of catalytic activity. A ) Effect of UBE2I on BrUTP incorporation. Day 13 meiotically incompetent oocytes were injected with water ( a , d , and g ), wild-type Ube2i mRNA ( b , e , and h ), or Ube2i C93S mRNA (
    Figure Legend Snippet: UBE2I stimulates BrUTP incorporation independent of catalytic activity. A ) Effect of UBE2I on BrUTP incorporation. Day 13 meiotically incompetent oocytes were injected with water ( a , d , and g ), wild-type Ube2i mRNA ( b , e , and h ), or Ube2i C93S mRNA (

    Techniques Used: Activity Assay, Injection

    Reorganization of UBE2I-containing bodies in response to α-amanitin. A ) Localization of UBE2I-containing bodies and sites of transcription. Day 13 incompetent oocytes were incubated for 12 h without ( a , d , g , and j ) or with 10 μg/ml (
    Figure Legend Snippet: Reorganization of UBE2I-containing bodies in response to α-amanitin. A ) Localization of UBE2I-containing bodies and sites of transcription. Day 13 incompetent oocytes were incubated for 12 h without ( a , d , g , and j ) or with 10 μg/ml (

    Techniques Used: Incubation

    Temporal pattern of Ube2i mRNA and UBE2I protein expression during oocyte growth and preimplantation development. A ) Ube2i transcript abundance was determined by quantitative RT-PCR and expressed relative to fully grown GV-intact oocytes. The results
    Figure Legend Snippet: Temporal pattern of Ube2i mRNA and UBE2I protein expression during oocyte growth and preimplantation development. A ) Ube2i transcript abundance was determined by quantitative RT-PCR and expressed relative to fully grown GV-intact oocytes. The results

    Techniques Used: Expressing, Quantitative RT-PCR

    Colocalization of UBE2I with SFRS2. Subcellular localization of UBE2I and SFRS2 in the nucleus of a Day 13 incompetent oocyte ( A ), a fully grown oocyte ( B ), and Hela S3 cells ( C ). a – d ) UBE2I ( a ), SFRS2 ( b ), DNA (DAPI) ( c ), and merged image ( d
    Figure Legend Snippet: Colocalization of UBE2I with SFRS2. Subcellular localization of UBE2I and SFRS2 in the nucleus of a Day 13 incompetent oocyte ( A ), a fully grown oocyte ( B ), and Hela S3 cells ( C ). a – d ) UBE2I ( a ), SFRS2 ( b ), DNA (DAPI) ( c ), and merged image ( d

    Techniques Used:

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    Article Snippet: .. Incubation with primary M2 anti-FLAG antibody (Sigma, no. F3165) was followed by incubation with secondary Tetramethyl Rhodamine 5 (and 6)-isothiocyanate (TRITC) conjugated goat-anti-mouse antibody (Jackson ImmunoResearch). .. qRT-PCR RNA was isolated from late 10,000 L4 worms per condition using the Trizol method. cDNA was synthesized using SuperScript III First-Strand Synthesis System (Invitrogen). qRT-PCR was performed with SsoAdvanced Univeral SYBR Green Supermix (Bio-Rad).

    Article Title: Effects of Combined Alcohol and Anti-HIV Drugs on Cellular Stress Responses in Primary Hepatocytes and Hepatic Stellate and Kupffer Cells
    Article Snippet: .. Coverslips were incubated with the anti-Nrf2 antibodies for 1 hour and then probed with a rhodamine TRITC fluorescent antibody (Jackson Immunoresearch, West Grove PA) for another hour. .. Filamentous actin double staining was performed using Alexa Fluor 488 conjugated phalloidin (Life Technologies, Grand Island, NY).

    Immunofluorescence:

    Article Title: Wt1 dictates the fate of fetal and adult Leydig cells during development in the mouse testis
    Article Snippet: .. Frozen cross-sections of testes 8 μm thick were fixed in 4% paraformaldehyde for 10 min. Immunofluorescence (IF) was performed using the FITC- or tetramethylrhodamine isothiocyanate (TRITC)-conjugated secondary antibodies (Jackson ImmunoResearch). .. Antibodies were obtained commercially as follows: Hsd3b1 (1:400, sc-30820; Santa Cruz Biotechnology), Ki-67 (1:1,000, ab15580; Abcam), P450scc (1:500, AB1244; Chemicon/Millipore), β-galactosidase (β-gal; 1:200, MP55976 ; MP Biomedical), and Vcam1 (1:400, AF643; R & D Systems).

    Confocal Microscopy:

    Article Title: Platelets Enhance Dendritic Cell Responses against Staphylococcus aureus through CD40-CD40L
    Article Snippet: .. The cells were then stained with TRITC-conjugated anti-IgG secondary antibody (Jackson ImmunoResearch, West Grove, PA) for 30 min at 4°C, washed at 600 × g for 5 min with DPBS−/− , resuspended in 10 μl of DPBS−/− , placed on coverslips (using Prolong Gold antifade reagent; Invitrogen; ), and observed under confocal microscopy to distinguish intracellular (FITC+ TRITC− ) from extracellular (FITC+ TRITC+ ) bacteria. .. Microscopy was performed using an Olympus FV1000 confocal microscope.

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  • 93
    Jackson Immuno donkey anti goat igg horseradish peroxidase conjugated secondary antibody
    100-fold less RABV-ED51-mBAFF is needed to induce comparable immunity in mice immunized with RABV. C57BL/6J mice were immunized i.m. with 10 3 or 10 5 ffu/mouse of RABV or RABV-ED51-mBAFF, or <t>PBS</t> as a negative control. Blood was collected on days 5, 7 and 10 post-immunization as a source of serum. Four three-fold serial dilutions of sera were analyzed by ELISA to determine anti-RABV G IgM (A and B) or <t>IgG</t> (C and D) antibody titers and presented as OD 490 of the reciprocal serial dilution. For comparison, sera from PBS-immunized mice were tested in parallel. Statistical difference in antibody titers by ELISA between RABV- and RABV-ED51-mBAFF-immunized mice was determined using an unpaired, two-tailed t test and data is presented as the mean ± SEM. *p≤0.05; **p ≤ 0.01; ***p≤0.001; ****p≤0.0001; N = 10 mice/group). (ffu = focus forming units; OD = optical density).
    Donkey Anti Goat Igg Horseradish Peroxidase Conjugated Secondary Antibody, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Jackson Immuno peroxidase conjugated donkey anti goat igg secondary antibody
    100-fold less RABV-ED51-mBAFF is needed to induce comparable immunity in mice immunized with RABV. C57BL/6J mice were immunized i.m. with 10 3 or 10 5 ffu/mouse of RABV or RABV-ED51-mBAFF, or <t>PBS</t> as a negative control. Blood was collected on days 5, 7 and 10 post-immunization as a source of serum. Four three-fold serial dilutions of sera were analyzed by ELISA to determine anti-RABV G IgM (A and B) or <t>IgG</t> (C and D) antibody titers and presented as OD 490 of the reciprocal serial dilution. For comparison, sera from PBS-immunized mice were tested in parallel. Statistical difference in antibody titers by ELISA between RABV- and RABV-ED51-mBAFF-immunized mice was determined using an unpaired, two-tailed t test and data is presented as the mean ± SEM. *p≤0.05; **p ≤ 0.01; ***p≤0.001; ****p≤0.0001; N = 10 mice/group). (ffu = focus forming units; OD = optical density).
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    Jackson Immuno peroxidase affinipure donkey anti human igg
    100-fold less RABV-ED51-mBAFF is needed to induce comparable immunity in mice immunized with RABV. C57BL/6J mice were immunized i.m. with 10 3 or 10 5 ffu/mouse of RABV or RABV-ED51-mBAFF, or <t>PBS</t> as a negative control. Blood was collected on days 5, 7 and 10 post-immunization as a source of serum. Four three-fold serial dilutions of sera were analyzed by ELISA to determine anti-RABV G IgM (A and B) or <t>IgG</t> (C and D) antibody titers and presented as OD 490 of the reciprocal serial dilution. For comparison, sera from PBS-immunized mice were tested in parallel. Statistical difference in antibody titers by ELISA between RABV- and RABV-ED51-mBAFF-immunized mice was determined using an unpaired, two-tailed t test and data is presented as the mean ± SEM. *p≤0.05; **p ≤ 0.01; ***p≤0.001; ****p≤0.0001; N = 10 mice/group). (ffu = focus forming units; OD = optical density).
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    Jackson Immuno hrp conjugated donkey anti goat igg antibody
    100-fold less RABV-ED51-mBAFF is needed to induce comparable immunity in mice immunized with RABV. C57BL/6J mice were immunized i.m. with 10 3 or 10 5 ffu/mouse of RABV or RABV-ED51-mBAFF, or <t>PBS</t> as a negative control. Blood was collected on days 5, 7 and 10 post-immunization as a source of serum. Four three-fold serial dilutions of sera were analyzed by ELISA to determine anti-RABV G IgM (A and B) or <t>IgG</t> (C and D) antibody titers and presented as OD 490 of the reciprocal serial dilution. For comparison, sera from PBS-immunized mice were tested in parallel. Statistical difference in antibody titers by ELISA between RABV- and RABV-ED51-mBAFF-immunized mice was determined using an unpaired, two-tailed t test and data is presented as the mean ± SEM. *p≤0.05; **p ≤ 0.01; ***p≤0.001; ****p≤0.0001; N = 10 mice/group). (ffu = focus forming units; OD = optical density).
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    Image Search Results


    100-fold less RABV-ED51-mBAFF is needed to induce comparable immunity in mice immunized with RABV. C57BL/6J mice were immunized i.m. with 10 3 or 10 5 ffu/mouse of RABV or RABV-ED51-mBAFF, or PBS as a negative control. Blood was collected on days 5, 7 and 10 post-immunization as a source of serum. Four three-fold serial dilutions of sera were analyzed by ELISA to determine anti-RABV G IgM (A and B) or IgG (C and D) antibody titers and presented as OD 490 of the reciprocal serial dilution. For comparison, sera from PBS-immunized mice were tested in parallel. Statistical difference in antibody titers by ELISA between RABV- and RABV-ED51-mBAFF-immunized mice was determined using an unpaired, two-tailed t test and data is presented as the mean ± SEM. *p≤0.05; **p ≤ 0.01; ***p≤0.001; ****p≤0.0001; N = 10 mice/group). (ffu = focus forming units; OD = optical density).

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Incorporating B cell activating factor (BAFF) into the membrane of rabies virus (RABV) particles improves the speed and magnitude of vaccine-induced antibody responses

    doi: 10.1371/journal.pntd.0007800

    Figure Lengend Snippet: 100-fold less RABV-ED51-mBAFF is needed to induce comparable immunity in mice immunized with RABV. C57BL/6J mice were immunized i.m. with 10 3 or 10 5 ffu/mouse of RABV or RABV-ED51-mBAFF, or PBS as a negative control. Blood was collected on days 5, 7 and 10 post-immunization as a source of serum. Four three-fold serial dilutions of sera were analyzed by ELISA to determine anti-RABV G IgM (A and B) or IgG (C and D) antibody titers and presented as OD 490 of the reciprocal serial dilution. For comparison, sera from PBS-immunized mice were tested in parallel. Statistical difference in antibody titers by ELISA between RABV- and RABV-ED51-mBAFF-immunized mice was determined using an unpaired, two-tailed t test and data is presented as the mean ± SEM. *p≤0.05; **p ≤ 0.01; ***p≤0.001; ****p≤0.0001; N = 10 mice/group). (ffu = focus forming units; OD = optical density).

    Article Snippet: Membrane was probed for one hour with polyclonal goat IgG anti-murine BAFF primary antibody (AF2106; R & D Systems) at a dilution of 1:2,000 in PBS-0.05% Tween-20 (PBS-T), washed 3 times with PBS-T, then incubated for one hour with donkey anti-goat IgG horseradish peroxidase-conjugated secondary antibody (Jackson Immuno) diluted 1:30,000 in PBS-T.

    Techniques: Mouse Assay, Negative Control, Enzyme-linked Immunosorbent Assay, Serial Dilution, Two Tailed Test