fluorescein labeled l pha  (Vector Laboratories)


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    Structured Review

    Vector Laboratories fluorescein labeled l pha
    CTD110.6 recognizes both O- GlcNAc and terminal β-GlcNAc on cell surface glycoproteins. A , N- ), of swainsonine. B , binding of <t>L-PHA</t> ( upper ) and mAb CTD110.6 ( lower ) to fixed cells. Lec2 and Lec8 cells were cultured with or without 5 μg/ml swainsonine ( SW ) for 4 days. Cells (4 × 10 5 ) were incubated with 20 μg/ml fluorescein-labeled L-PHA or 40 μg/ml CTD110.6 on ice for 20 min followed by 10 μg/ml Cy5-conjugated anti-mouse IgM antibody. Gray profiles , secondary Abs; thin line , control cells; bold line , swainsonine-treated cells. Representative results are from two independent experiments.
    Fluorescein Labeled L Pha, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fluorescein labeled l pha/product/Vector Laboratories
    Average 93 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    fluorescein labeled l pha - by Bioz Stars, 2022-09
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    Images

    1) Product Images from "Antibodies That Detect O-Linked β-d-N-Acetylglucosamine on the Extracellular Domain of Cell Surface Glycoproteins *"

    Article Title: Antibodies That Detect O-Linked β-d-N-Acetylglucosamine on the Extracellular Domain of Cell Surface Glycoproteins *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M113.492512

    CTD110.6 recognizes both O- GlcNAc and terminal β-GlcNAc on cell surface glycoproteins. A , N- ), of swainsonine. B , binding of L-PHA ( upper ) and mAb CTD110.6 ( lower ) to fixed cells. Lec2 and Lec8 cells were cultured with or without 5 μg/ml swainsonine ( SW ) for 4 days. Cells (4 × 10 5 ) were incubated with 20 μg/ml fluorescein-labeled L-PHA or 40 μg/ml CTD110.6 on ice for 20 min followed by 10 μg/ml Cy5-conjugated anti-mouse IgM antibody. Gray profiles , secondary Abs; thin line , control cells; bold line , swainsonine-treated cells. Representative results are from two independent experiments.
    Figure Legend Snippet: CTD110.6 recognizes both O- GlcNAc and terminal β-GlcNAc on cell surface glycoproteins. A , N- ), of swainsonine. B , binding of L-PHA ( upper ) and mAb CTD110.6 ( lower ) to fixed cells. Lec2 and Lec8 cells were cultured with or without 5 μg/ml swainsonine ( SW ) for 4 days. Cells (4 × 10 5 ) were incubated with 20 μg/ml fluorescein-labeled L-PHA or 40 μg/ml CTD110.6 on ice for 20 min followed by 10 μg/ml Cy5-conjugated anti-mouse IgM antibody. Gray profiles , secondary Abs; thin line , control cells; bold line , swainsonine-treated cells. Representative results are from two independent experiments.

    Techniques Used: Binding Assay, Cell Culture, Incubation, Labeling

    2) Product Images from "A testis-specific regulator of complex and hybrid N-glycan synthesis"

    Article Title: A testis-specific regulator of complex and hybrid N-glycan synthesis

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201004102

    GlcNAcT-I-KDEL localized mainly in the ER is active. Lec1 cells stably expressing empty vector, GlcNAcT-I-HA-KDEL, or GlcNAcT-I-HA were analyzed for (A) Con A resistance and (B) L-PHA-FITC binding by flow cytometry. (C) GlcNAcT-I and β4GalT activities in transfectant lysates were determined. Bars represent range ( n = 2). (D) HeLa cells transiently expressing GlcNAcT-I-HA-KDEL were fixed and immunofluorescently labeled for GlcNAcT-I-HA-KDEL (red) and endogenous ManII (green). Bars, 10 µm.
    Figure Legend Snippet: GlcNAcT-I-KDEL localized mainly in the ER is active. Lec1 cells stably expressing empty vector, GlcNAcT-I-HA-KDEL, or GlcNAcT-I-HA were analyzed for (A) Con A resistance and (B) L-PHA-FITC binding by flow cytometry. (C) GlcNAcT-I and β4GalT activities in transfectant lysates were determined. Bars represent range ( n = 2). (D) HeLa cells transiently expressing GlcNAcT-I-HA-KDEL were fixed and immunofluorescently labeled for GlcNAcT-I-HA-KDEL (red) and endogenous ManII (green). Bars, 10 µm.

    Techniques Used: Stable Transfection, Expressing, Plasmid Preparation, Binding Assay, Flow Cytometry, Cytometry, Transfection, Labeling

    3) Product Images from "Overexpression of CD47 is associated with brain overgrowth in 16p11.2 deletion syndrome"

    Article Title: Overexpression of CD47 is associated with brain overgrowth in 16p11.2 deletion syndrome

    Journal: bioRxiv

    doi: 10.1101/808022

    Calreticulin, a pro-phagocytic signal, is upregulated in 16p11.2 deletion NPCs and OPCs. A. Representative histograms show the mean fluorescence intensities (MFI) of cell-surface expression of Calreticulin (CRT) in NPCs derived from control, 16p_del, and 16p_dup lines. B. Quantification of CRT MFI of cell-surface expression of CRT in the 16p_del NPCs relative to 16p_dup and control NPCs. C. Quantification of Phaseolus Vulgaris Leucoagglutinin (PHA-L), indicative of asialoglycan binding sites for CRT, in NPCs in 16p_deletion lines relative to control and 16p_duplication lines. D. Representative histograms show the MFI of cell-surface expression of CRT in OPCs derived from control, 16p_del, and 16p_dup lines. E. Quantification of CRT MFI of cell-surface expression of CRT in the 16p_del OPCs relative to 16p_dup and control OPCs. F. Quantification of PHA-L in OPCs in 16p_deletion lines relative to control and 16p_duplication lines. n=2 biological replicates per cell line (16p_dup, n=2; control, n=3; 16p_del, n=4 cell lines). All data are mean ± SE; P values were determined by one-way ANOVA followed by post-hoc Tukey HSD Test.
    Figure Legend Snippet: Calreticulin, a pro-phagocytic signal, is upregulated in 16p11.2 deletion NPCs and OPCs. A. Representative histograms show the mean fluorescence intensities (MFI) of cell-surface expression of Calreticulin (CRT) in NPCs derived from control, 16p_del, and 16p_dup lines. B. Quantification of CRT MFI of cell-surface expression of CRT in the 16p_del NPCs relative to 16p_dup and control NPCs. C. Quantification of Phaseolus Vulgaris Leucoagglutinin (PHA-L), indicative of asialoglycan binding sites for CRT, in NPCs in 16p_deletion lines relative to control and 16p_duplication lines. D. Representative histograms show the MFI of cell-surface expression of CRT in OPCs derived from control, 16p_del, and 16p_dup lines. E. Quantification of CRT MFI of cell-surface expression of CRT in the 16p_del OPCs relative to 16p_dup and control OPCs. F. Quantification of PHA-L in OPCs in 16p_deletion lines relative to control and 16p_duplication lines. n=2 biological replicates per cell line (16p_dup, n=2; control, n=3; 16p_del, n=4 cell lines). All data are mean ± SE; P values were determined by one-way ANOVA followed by post-hoc Tukey HSD Test.

    Techniques Used: Fluorescence, Expressing, Derivative Assay, Binding Assay

    4) Product Images from "The activation of PPARγ enhances Treg responses through up-regulating CD36/CPT1-mediated fatty acid oxidation and subsequent N-glycan branching of TβRII/IL-2Rα"

    Article Title: The activation of PPARγ enhances Treg responses through up-regulating CD36/CPT1-mediated fatty acid oxidation and subsequent N-glycan branching of TβRII/IL-2Rα

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/s12964-022-00849-9

    PFK participates in the effects of PPARγ agonists through associating FAO with UDP-GlcNAc/glycosylation axis. a – d The naïve CD4 + T cells were transfected with siCD36 (50, 100, 150, 200 nM) or siCPT1 (50, 100, 150, 200 nM), and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL) for 48 h. The mRNA levels of GFPT and PFK were measured by Q-PCR ( a , b ), and the activity of PFK and GFPT was determined ( c , d ). e The naïve CD4 + T cells were prepared and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (10, 30 μM), 15d-PGJ2 (3, 10 μM) as well as morin (10, 30 μM) for 48 h. The activity of PFK and GFPT was determined. f The naïve CD4 + T cells were transfected with siPFK, and the mRNA level of PFK was measured by Q-PCR. g – i The naïve CD4 + T cells were transfected with siPFK, and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (30 μM), 15d-PGJ2 (10 μM) as well as morin (30 μM) for 48 h. The intracellular level of UDP-GlcNAc was determined ( g ), and the level of N-linked glycosylation was examined by flow cytometry using FITC-L-PHA ( h , i ). j – l The naïve CD4 + T cells were transfected with siCD36 or siCPT1, and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (30 μM), 15d-PGJ2 (10 μM) as well as morin (30 μM) for 48 h. The activity of PFK ( j ) and the intracellular level of UDP-GlcNAc ( k ) were determined. The level of N-linked glycosylation was examined by flow cytometry using FITC-L-PHA ( l ). Data were presented as the means ± S.E.M. of three independent experiments (n = 3). ## P
    Figure Legend Snippet: PFK participates in the effects of PPARγ agonists through associating FAO with UDP-GlcNAc/glycosylation axis. a – d The naïve CD4 + T cells were transfected with siCD36 (50, 100, 150, 200 nM) or siCPT1 (50, 100, 150, 200 nM), and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL) for 48 h. The mRNA levels of GFPT and PFK were measured by Q-PCR ( a , b ), and the activity of PFK and GFPT was determined ( c , d ). e The naïve CD4 + T cells were prepared and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (10, 30 μM), 15d-PGJ2 (3, 10 μM) as well as morin (10, 30 μM) for 48 h. The activity of PFK and GFPT was determined. f The naïve CD4 + T cells were transfected with siPFK, and the mRNA level of PFK was measured by Q-PCR. g – i The naïve CD4 + T cells were transfected with siPFK, and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (30 μM), 15d-PGJ2 (10 μM) as well as morin (30 μM) for 48 h. The intracellular level of UDP-GlcNAc was determined ( g ), and the level of N-linked glycosylation was examined by flow cytometry using FITC-L-PHA ( h , i ). j – l The naïve CD4 + T cells were transfected with siCD36 or siCPT1, and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (30 μM), 15d-PGJ2 (10 μM) as well as morin (30 μM) for 48 h. The activity of PFK ( j ) and the intracellular level of UDP-GlcNAc ( k ) were determined. The level of N-linked glycosylation was examined by flow cytometry using FITC-L-PHA ( l ). Data were presented as the means ± S.E.M. of three independent experiments (n = 3). ## P

    Techniques Used: Transfection, Polymerase Chain Reaction, Activity Assay, Flow Cytometry

    PPARγ agonists increase the level of UDP-GlcNAc and subsequent N-glycan branching during Treg differentiation. a The process of hexosamine biosynthesis pathway and the N-glycan biosynthesis was shown. b – d The naïve CD4 + T cells were prepared and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (10, 30 μM), 15d-PGJ2 (3, 10 μM) as well as morin (10, 30 μM) for 48 h. The intracellular level of UDP-GlcNAc was detected ( b ), the level of N-linked glycosylation was examined by flow cytometry using FITC-L-PHA ( c ), and the mRNA expression levels of Mgat1, Mgat2, Mgat4 and Mgat5 ( d ) were measured by Q-PCR. ## P
    Figure Legend Snippet: PPARγ agonists increase the level of UDP-GlcNAc and subsequent N-glycan branching during Treg differentiation. a The process of hexosamine biosynthesis pathway and the N-glycan biosynthesis was shown. b – d The naïve CD4 + T cells were prepared and treated with anti-CD3/CD28 in the presence or absence of TGF-β (5 ng/mL), rosiglitazone (10, 30 μM), 15d-PGJ2 (3, 10 μM) as well as morin (10, 30 μM) for 48 h. The intracellular level of UDP-GlcNAc was detected ( b ), the level of N-linked glycosylation was examined by flow cytometry using FITC-L-PHA ( c ), and the mRNA expression levels of Mgat1, Mgat2, Mgat4 and Mgat5 ( d ) were measured by Q-PCR. ## P

    Techniques Used: Flow Cytometry, Expressing, Polymerase Chain Reaction

    5) Product Images from "Hybrid- and complex-type N-glycans are not essential for Newcastle disease virus infection and fusion of host cells"

    Article Title: Hybrid- and complex-type N-glycans are not essential for Newcastle disease virus infection and fusion of host cells

    Journal: Glycobiology

    doi: 10.1093/glycob/cwr146

    Characterization of Lec1 cells phenotype. CHO-K1 ( B ) and Lec1 ( A ) cells were fixed in 2% paraformaldehyde, blocked in TBS containing 10% goat serum, and then incubated with FITC-labeled lectin L-PHA, with nuclei counterstained with DAPI. Cells were visualized under Leica confocal fluorescence microscope (TCS SP2). CHO-K1 (red line) and Lec1 cells (black line) were incubated with FITC-labeled L-PHA for 45 min at 4°C and analyzed by flow cytometry. Cells analyzed in the absence of FITC-labeled L-PHA are shown in the filled light-gray trace ( C ). CHO-K1 ( E ) and Lec1 ( D ) cells were fixed in 2% paraformaldehyde, blocked in TBS containing 10% goat serum, and then incubated with DIG-labeled lectin GNA at 37°C. After 45 min incubation, cells were incubated with AP-conjugated anti-DIG for 45 min at 37°C and then treated with NBT/BCIP for visualization under Leical fluorescence microscope. CHO-K1 (red line) and Lec1 cells (black line) were incubated with the DIG-labeled lectin GNA for 45 min at 4°C and incubated with FITC-conjugated anti-DIG for 45 min at 37°C and then analyzed by flow cytometry. Cells analyzed in the absence of lectin are shown in the filled light-gray trace ( F ).
    Figure Legend Snippet: Characterization of Lec1 cells phenotype. CHO-K1 ( B ) and Lec1 ( A ) cells were fixed in 2% paraformaldehyde, blocked in TBS containing 10% goat serum, and then incubated with FITC-labeled lectin L-PHA, with nuclei counterstained with DAPI. Cells were visualized under Leica confocal fluorescence microscope (TCS SP2). CHO-K1 (red line) and Lec1 cells (black line) were incubated with FITC-labeled L-PHA for 45 min at 4°C and analyzed by flow cytometry. Cells analyzed in the absence of FITC-labeled L-PHA are shown in the filled light-gray trace ( C ). CHO-K1 ( E ) and Lec1 ( D ) cells were fixed in 2% paraformaldehyde, blocked in TBS containing 10% goat serum, and then incubated with DIG-labeled lectin GNA at 37°C. After 45 min incubation, cells were incubated with AP-conjugated anti-DIG for 45 min at 37°C and then treated with NBT/BCIP for visualization under Leical fluorescence microscope. CHO-K1 (red line) and Lec1 cells (black line) were incubated with the DIG-labeled lectin GNA for 45 min at 4°C and incubated with FITC-conjugated anti-DIG for 45 min at 37°C and then analyzed by flow cytometry. Cells analyzed in the absence of lectin are shown in the filled light-gray trace ( F ).

    Techniques Used: Incubation, Labeling, Fluorescence, Microscopy, Flow Cytometry, Cytometry

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    Vector Laboratories fluorescein labeled l pha
    CTD110.6 recognizes both O- GlcNAc and terminal β-GlcNAc on cell surface glycoproteins. A , N- ), of swainsonine. B , binding of <t>L-PHA</t> ( upper ) and mAb CTD110.6 ( lower ) to fixed cells. Lec2 and Lec8 cells were cultured with or without 5 μg/ml swainsonine ( SW ) for 4 days. Cells (4 × 10 5 ) were incubated with 20 μg/ml fluorescein-labeled L-PHA or 40 μg/ml CTD110.6 on ice for 20 min followed by 10 μg/ml Cy5-conjugated anti-mouse IgM antibody. Gray profiles , secondary Abs; thin line , control cells; bold line , swainsonine-treated cells. Representative results are from two independent experiments.
    Fluorescein Labeled L Pha, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fluorescein labeled l pha/product/Vector Laboratories
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    95
    Vector Laboratories phaseolus vulgaris leucoagglutinin
    High glucose-stimulated UT-A1 sialylation is blocked by a PKC inhibitor. (A and B) UT-A1-MDCK cells were preincubated with 5.5 mM glucose medium for 24 hours and then incubated with DMEM containing 5.5 or 25 mM glucose or 25 mM L-glucose for another 24 hours. Cells were lysed in RIPA buffer, equal amounts of total cell lysates were used for GNL and SNA lectin pull-down assay, and UT-A1 protein expression was analyzed by Western blot. (A) Total or (B) lectin-precipitated UT-A1 was examined by Western blot. (C) UT-A1 MDCK cells were treated as above, except for that one group had a 2-hour pretreatment with 10 µ M chelerythrine (Chel). The bar graph shows band densities as fold of control (means±SDs; n =3). Ctrl, control; DSL, datura stramonium lectin; IB, immunoblot; PHA, <t>phaseolus</t> <t>vulgaris</t> <t>leucoagglutinin;</t> Tomato, lycopersicum esculentum lectin; WGA, wheat germ agglutinin. * P
    Phaseolus Vulgaris Leucoagglutinin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Vector Laboratories biotinylated pha l
    Loss of MGAT4A activity does not affect GlcNAc-β-1,6 branching. A , biosynthetic pathway of tetraantennary branched N -glycans. Binding sites of <t>PHA-L</t> are indicated. B , analysis of PHA-L binding to the Consortium of Functional Glycomics ( CFG ) glycan array v. 5.0 indicates a preference for GlcNAc-β-1,6 branched products. The graph shows the data for 100 μg/ml <t>biotinylated-PHA-L</t> (Vector Laboratories) binding to glycans bearing only β-1,4 branching (β -(1,4) ) and the corresponding β-1,6 glycans (β -(1,6) ). Data are given as fluorescence intensity. C , knockdown of MGAT4A in MCF-7 cells shows no significant change in MGAT4B or MGAT5 mRNA levels by qRT-PCR. The graph shows -fold change mRNA expression compared with NTC. n = 3 biological replicates. Error bars represent the S.D. D , fluorescence microscopy of PHA-L stained MGAT4A-KD and NTC cells. No significant loss of PHA-L binding were observed. Images shown are representative of two biological replicates, five images per replicate.
    Biotinylated Pha L, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    CTD110.6 recognizes both O- GlcNAc and terminal β-GlcNAc on cell surface glycoproteins. A , N- ), of swainsonine. B , binding of L-PHA ( upper ) and mAb CTD110.6 ( lower ) to fixed cells. Lec2 and Lec8 cells were cultured with or without 5 μg/ml swainsonine ( SW ) for 4 days. Cells (4 × 10 5 ) were incubated with 20 μg/ml fluorescein-labeled L-PHA or 40 μg/ml CTD110.6 on ice for 20 min followed by 10 μg/ml Cy5-conjugated anti-mouse IgM antibody. Gray profiles , secondary Abs; thin line , control cells; bold line , swainsonine-treated cells. Representative results are from two independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Antibodies That Detect O-Linked β-d-N-Acetylglucosamine on the Extracellular Domain of Cell Surface Glycoproteins *

    doi: 10.1074/jbc.M113.492512

    Figure Lengend Snippet: CTD110.6 recognizes both O- GlcNAc and terminal β-GlcNAc on cell surface glycoproteins. A , N- ), of swainsonine. B , binding of L-PHA ( upper ) and mAb CTD110.6 ( lower ) to fixed cells. Lec2 and Lec8 cells were cultured with or without 5 μg/ml swainsonine ( SW ) for 4 days. Cells (4 × 10 5 ) were incubated with 20 μg/ml fluorescein-labeled L-PHA or 40 μg/ml CTD110.6 on ice for 20 min followed by 10 μg/ml Cy5-conjugated anti-mouse IgM antibody. Gray profiles , secondary Abs; thin line , control cells; bold line , swainsonine-treated cells. Representative results are from two independent experiments.

    Article Snippet: Washed cells were incubated with 50 μl of binding buffer containing 1 μg of fluorescein-labeled L-PHA ( Phaseolus vulgaris leukoagglutinin; Vector, Burlingame, CA) or 2 μg of CTD110.6 mAb, followed by incubation with 50 μl of binding buffer containing 0.5 μg of Cy5-conjugated anti-mouse IgM.

    Techniques: Binding Assay, Cell Culture, Incubation, Labeling

    High glucose-stimulated UT-A1 sialylation is blocked by a PKC inhibitor. (A and B) UT-A1-MDCK cells were preincubated with 5.5 mM glucose medium for 24 hours and then incubated with DMEM containing 5.5 or 25 mM glucose or 25 mM L-glucose for another 24 hours. Cells were lysed in RIPA buffer, equal amounts of total cell lysates were used for GNL and SNA lectin pull-down assay, and UT-A1 protein expression was analyzed by Western blot. (A) Total or (B) lectin-precipitated UT-A1 was examined by Western blot. (C) UT-A1 MDCK cells were treated as above, except for that one group had a 2-hour pretreatment with 10 µ M chelerythrine (Chel). The bar graph shows band densities as fold of control (means±SDs; n =3). Ctrl, control; DSL, datura stramonium lectin; IB, immunoblot; PHA, phaseolus vulgaris leucoagglutinin; Tomato, lycopersicum esculentum lectin; WGA, wheat germ agglutinin. * P

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: Activation of Protein Kinase C-α and Src Kinase Increases Urea Transporter A1 α-2, 6 Sialylation

    doi: 10.1681/ASN.2014010026

    Figure Lengend Snippet: High glucose-stimulated UT-A1 sialylation is blocked by a PKC inhibitor. (A and B) UT-A1-MDCK cells were preincubated with 5.5 mM glucose medium for 24 hours and then incubated with DMEM containing 5.5 or 25 mM glucose or 25 mM L-glucose for another 24 hours. Cells were lysed in RIPA buffer, equal amounts of total cell lysates were used for GNL and SNA lectin pull-down assay, and UT-A1 protein expression was analyzed by Western blot. (A) Total or (B) lectin-precipitated UT-A1 was examined by Western blot. (C) UT-A1 MDCK cells were treated as above, except for that one group had a 2-hour pretreatment with 10 µ M chelerythrine (Chel). The bar graph shows band densities as fold of control (means±SDs; n =3). Ctrl, control; DSL, datura stramonium lectin; IB, immunoblot; PHA, phaseolus vulgaris leucoagglutinin; Tomato, lycopersicum esculentum lectin; WGA, wheat germ agglutinin. * P

    Article Snippet: Agarose-bound concanavalin A, wheat germ agglutinin, SNA, GNL, datura stramonium lectin, phaseolus vulgaris leucoagglutinin, and Tomato lectin (Lycopersicum esculentum lectin) were purchased from Vector Laboratories (Burlingame, CA).

    Techniques: Incubation, Pull Down Assay, Expressing, Western Blot, Whole Genome Amplification

    PKC activator increases UT-A1 glycan sialylation. (A) UT-A1 MDCK cells were treated with 2 µ M PDBu for 24 hours. The cells were lysed in RIPA buffer, equal amounts of total lysates were incubated with agarose-conjugated lectins, and then, they were immunoblotted with antibody to UT-A1. (B) Kidney IMCD suspensions prepared from Sprague–Dawley rats were treated with 2 µ M PDBu for 6 hours. The cell membrane lipid rafts were prepared by a 5%–40% sucrose gradient ultracentrifugation. Fractions 2–5 were collected for lectin pull-down assay with different lectins. The lectin-precipitated samples were then analyzed for UT-A1 protein expression by Western blot. The bar graph shows band densities as fold of control (means±SDs; n =3). Con A, concanavalin A; Ctrl, control; DSL, datura stramonium lectin; IB, immunoblot; PHA, phaseolus vulgaris leucoagglutinin; Tomato, lycopersicum esculentum lectin; WGA, wheat germ agglutinin. ** P

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: Activation of Protein Kinase C-α and Src Kinase Increases Urea Transporter A1 α-2, 6 Sialylation

    doi: 10.1681/ASN.2014010026

    Figure Lengend Snippet: PKC activator increases UT-A1 glycan sialylation. (A) UT-A1 MDCK cells were treated with 2 µ M PDBu for 24 hours. The cells were lysed in RIPA buffer, equal amounts of total lysates were incubated with agarose-conjugated lectins, and then, they were immunoblotted with antibody to UT-A1. (B) Kidney IMCD suspensions prepared from Sprague–Dawley rats were treated with 2 µ M PDBu for 6 hours. The cell membrane lipid rafts were prepared by a 5%–40% sucrose gradient ultracentrifugation. Fractions 2–5 were collected for lectin pull-down assay with different lectins. The lectin-precipitated samples were then analyzed for UT-A1 protein expression by Western blot. The bar graph shows band densities as fold of control (means±SDs; n =3). Con A, concanavalin A; Ctrl, control; DSL, datura stramonium lectin; IB, immunoblot; PHA, phaseolus vulgaris leucoagglutinin; Tomato, lycopersicum esculentum lectin; WGA, wheat germ agglutinin. ** P

    Article Snippet: Agarose-bound concanavalin A, wheat germ agglutinin, SNA, GNL, datura stramonium lectin, phaseolus vulgaris leucoagglutinin, and Tomato lectin (Lycopersicum esculentum lectin) were purchased from Vector Laboratories (Burlingame, CA).

    Techniques: Incubation, Pull Down Assay, Expressing, Western Blot, Whole Genome Amplification

    Camera-lucida reconstruction of axon 4-2 labeled by PHA-L anterogradely transported from TEav (case 4) to the ventrocaudal striatum and the dorsolateral aspect of the deep amygdaloid nuclei. Thick lines represent the main axonal trunk and collaterals, and thin lines represent terminal arbors. Numbers indicate serial numbers of individual sections; smaller numbers are posterior. This axon was serially reconstructed through 103 sections (section thickness, 40 μm). Ten terminal arbors ( ar1–ar10 ) were drawn; each arbor is presented with a range of sections ( numbers in parentheses ), from which the arbor was completely reconstructed. Sections in which the axon gives off major collaterals are indicated by section numbers and arrows . Dashed lines (with section numbers ) indicate borders between different structures. The global locations of terminal arbors are illustrated in low-magnification camera-lucida drawings of selected sections, where terminal clusters containing individual arbors are circumscribed by shaded ellipses or circles . Double lines indicate the incomplete portion of the axon. The main axonal trunk in the white matter was followed to the injection site. AB , Accessory basal nucleus; CE , central nucleus; L , lateral nucleus; LB , lateral basal nucleus; MB , medial basal nucleus; Amy , amygdala; Cd , caudate nucleus; Cl , claustrum; GPe , external globus pallidus; H , hippocampus; LV , lateral ventricle; Put , putamen; WM , white matter; rh , rhinal sulcus. Orientation: M , medial; L , lateral; D , dorsal; V , ventral. The same conventions and abbreviations are used for illustrating other reconstructed axons unless otherwise noted.

    Journal: The Journal of Neuroscience

    Article Title: Organization of Corticostriatal and Corticoamygdalar Projections Arising from the Anterior Inferotemporal Area TE of the Macaque Monkey: A Phaseolus vulgaris Leucoagglutinin Study

    doi: 10.1523/JNEUROSCI.17-20-07902.1997

    Figure Lengend Snippet: Camera-lucida reconstruction of axon 4-2 labeled by PHA-L anterogradely transported from TEav (case 4) to the ventrocaudal striatum and the dorsolateral aspect of the deep amygdaloid nuclei. Thick lines represent the main axonal trunk and collaterals, and thin lines represent terminal arbors. Numbers indicate serial numbers of individual sections; smaller numbers are posterior. This axon was serially reconstructed through 103 sections (section thickness, 40 μm). Ten terminal arbors ( ar1–ar10 ) were drawn; each arbor is presented with a range of sections ( numbers in parentheses ), from which the arbor was completely reconstructed. Sections in which the axon gives off major collaterals are indicated by section numbers and arrows . Dashed lines (with section numbers ) indicate borders between different structures. The global locations of terminal arbors are illustrated in low-magnification camera-lucida drawings of selected sections, where terminal clusters containing individual arbors are circumscribed by shaded ellipses or circles . Double lines indicate the incomplete portion of the axon. The main axonal trunk in the white matter was followed to the injection site. AB , Accessory basal nucleus; CE , central nucleus; L , lateral nucleus; LB , lateral basal nucleus; MB , medial basal nucleus; Amy , amygdala; Cd , caudate nucleus; Cl , claustrum; GPe , external globus pallidus; H , hippocampus; LV , lateral ventricle; Put , putamen; WM , white matter; rh , rhinal sulcus. Orientation: M , medial; L , lateral; D , dorsal; V , ventral. The same conventions and abbreviations are used for illustrating other reconstructed axons unless otherwise noted.

    Article Snippet: PHA-L (2.5%; Vector Laboratories, Burlingame, CA) was injected iontophoretically (Midgard precision current source; Stoelting), according to the procedure recommended by with some modifications ( ).

    Techniques: Labeling, Injection

    Camera-lucida drawings of 13 coronal section outlines showing the distribution of PHA-L-labeled terminals in the ventrocaudal striatum (composed of the tail of the caudate nucleus and adjacent ventral putamen), the ventral striatum (composed of the nucleus accumbens, olfactory tubercle, and adjacent ventral part of the head of the caudate nucleus and ventral putamen), and the remaining parts of the striatum after TEav injection (case 3). Numbers indicate serial numbers of individual sections (40 μm thick); bottom section numbers indicate posterior sections. Section outlines are illustrated caudorostrally ( A–N ) at equal intervals (1.2 mm). Note the absence of labeled terminals at the level of the anterior commissure ( E, F ). AC , Anterior commissure; Cd , caudate nucleus; IC , internal capsule; NA , nucleus accumbens; OT , olfactory tubercle; Put , putamen.

    Journal: The Journal of Neuroscience

    Article Title: Organization of Corticostriatal and Corticoamygdalar Projections Arising from the Anterior Inferotemporal Area TE of the Macaque Monkey: A Phaseolus vulgaris Leucoagglutinin Study

    doi: 10.1523/JNEUROSCI.17-20-07902.1997

    Figure Lengend Snippet: Camera-lucida drawings of 13 coronal section outlines showing the distribution of PHA-L-labeled terminals in the ventrocaudal striatum (composed of the tail of the caudate nucleus and adjacent ventral putamen), the ventral striatum (composed of the nucleus accumbens, olfactory tubercle, and adjacent ventral part of the head of the caudate nucleus and ventral putamen), and the remaining parts of the striatum after TEav injection (case 3). Numbers indicate serial numbers of individual sections (40 μm thick); bottom section numbers indicate posterior sections. Section outlines are illustrated caudorostrally ( A–N ) at equal intervals (1.2 mm). Note the absence of labeled terminals at the level of the anterior commissure ( E, F ). AC , Anterior commissure; Cd , caudate nucleus; IC , internal capsule; NA , nucleus accumbens; OT , olfactory tubercle; Put , putamen.

    Article Snippet: PHA-L (2.5%; Vector Laboratories, Burlingame, CA) was injected iontophoretically (Midgard precision current source; Stoelting), according to the procedure recommended by with some modifications ( ).

    Techniques: Labeling, Injection

    Camera-lucida drawings of 8 coronal section outlines showing the distribution of PHA-L-labeled terminals in the amygdala after TEav injection (case 3). Numbers indicate serial numbers of individual sections (40 μm thick); bottom section numbers indicate posterior sections. Section outlines are illustrated caudorostrally ( A–H ) at equal intervals (0.6 mm). AAA , Anterior amygdaloid area; AB , accessory basal nucleus; CE , central nucleus; COa , anterior cortical nucleus; En , endopiriform nucleus; L , lateral nucleus; LB , lateral basal nucleus; M , medial nucleus; MB , medial basal nucleus; NLOT , nucleus of the lateral olfactory tract; PAC , periamygdaloid cortex; Pir , piriform cortex; DAN , deep amygdaloid nuclei; Cd , caudate nucleus; Cl , claustrum; EC , entorhinal cortex; H , hippocampus; LV , lateral ventricle; Put , putamen; rh , rhinal sulcus.

    Journal: The Journal of Neuroscience

    Article Title: Organization of Corticostriatal and Corticoamygdalar Projections Arising from the Anterior Inferotemporal Area TE of the Macaque Monkey: A Phaseolus vulgaris Leucoagglutinin Study

    doi: 10.1523/JNEUROSCI.17-20-07902.1997

    Figure Lengend Snippet: Camera-lucida drawings of 8 coronal section outlines showing the distribution of PHA-L-labeled terminals in the amygdala after TEav injection (case 3). Numbers indicate serial numbers of individual sections (40 μm thick); bottom section numbers indicate posterior sections. Section outlines are illustrated caudorostrally ( A–H ) at equal intervals (0.6 mm). AAA , Anterior amygdaloid area; AB , accessory basal nucleus; CE , central nucleus; COa , anterior cortical nucleus; En , endopiriform nucleus; L , lateral nucleus; LB , lateral basal nucleus; M , medial nucleus; MB , medial basal nucleus; NLOT , nucleus of the lateral olfactory tract; PAC , periamygdaloid cortex; Pir , piriform cortex; DAN , deep amygdaloid nuclei; Cd , caudate nucleus; Cl , claustrum; EC , entorhinal cortex; H , hippocampus; LV , lateral ventricle; Put , putamen; rh , rhinal sulcus.

    Article Snippet: PHA-L (2.5%; Vector Laboratories, Burlingame, CA) was injected iontophoretically (Midgard precision current source; Stoelting), according to the procedure recommended by with some modifications ( ).

    Techniques: Labeling, Injection

    Loss of MGAT4A activity does not affect GlcNAc-β-1,6 branching. A , biosynthetic pathway of tetraantennary branched N -glycans. Binding sites of PHA-L are indicated. B , analysis of PHA-L binding to the Consortium of Functional Glycomics ( CFG ) glycan array v. 5.0 indicates a preference for GlcNAc-β-1,6 branched products. The graph shows the data for 100 μg/ml biotinylated-PHA-L (Vector Laboratories) binding to glycans bearing only β-1,4 branching (β -(1,4) ) and the corresponding β-1,6 glycans (β -(1,6) ). Data are given as fluorescence intensity. C , knockdown of MGAT4A in MCF-7 cells shows no significant change in MGAT4B or MGAT5 mRNA levels by qRT-PCR. The graph shows -fold change mRNA expression compared with NTC. n = 3 biological replicates. Error bars represent the S.D. D , fluorescence microscopy of PHA-L stained MGAT4A-KD and NTC cells. No significant loss of PHA-L binding were observed. Images shown are representative of two biological replicates, five images per replicate.

    Journal: The Journal of Biological Chemistry

    Article Title: MicroRNA-424 Predicts a Role for β-1,4 Branched Glycosylation in Cell Cycle Progression *

    doi: 10.1074/jbc.M115.672220

    Figure Lengend Snippet: Loss of MGAT4A activity does not affect GlcNAc-β-1,6 branching. A , biosynthetic pathway of tetraantennary branched N -glycans. Binding sites of PHA-L are indicated. B , analysis of PHA-L binding to the Consortium of Functional Glycomics ( CFG ) glycan array v. 5.0 indicates a preference for GlcNAc-β-1,6 branched products. The graph shows the data for 100 μg/ml biotinylated-PHA-L (Vector Laboratories) binding to glycans bearing only β-1,4 branching (β -(1,4) ) and the corresponding β-1,6 glycans (β -(1,6) ). Data are given as fluorescence intensity. C , knockdown of MGAT4A in MCF-7 cells shows no significant change in MGAT4B or MGAT5 mRNA levels by qRT-PCR. The graph shows -fold change mRNA expression compared with NTC. n = 3 biological replicates. Error bars represent the S.D. D , fluorescence microscopy of PHA-L stained MGAT4A-KD and NTC cells. No significant loss of PHA-L binding were observed. Images shown are representative of two biological replicates, five images per replicate.

    Article Snippet: Samples were then blocked with 5% bovine serum albumin in HBSS (2 ml, 30 min) and incubated with primary antibodies or lectins diluted in HBSS as follows for 1 h at room temperature: α-galectin-3 (1:250), α-E-cadherin (1:500), biotinylated-PHA-L (1:500, Vector Laboratories).

    Techniques: Activity Assay, Binding Assay, Functional Assay, Plasmid Preparation, Fluorescence, Quantitative RT-PCR, Expressing, Microscopy, Staining