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biotinylated lectins  (Vector Laboratories)


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    Vector Laboratories biotinylated lectins
    Biotinylated Lectins, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    Vector Laboratories biotinylated lectins
    Biotinylated Lectins, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories biotinylated lectin con a
    The values of relative reactivities of serum IgG N-glycans with * PHA-L, Con A, and PHA-L/Con A ratio. The results are expressed in absorbance units (AUs) as a median with an interquartile range (Q1–Q3). Serum IgG glycan reactivities with <t>lectin</t> were examined by lectin-ELISA and expressed in AU. Significant differences versus groups: 1 with advanced endometriosis (AE), 2 non-endometriosis (NE), and control—a group of healthy women. Con A—relative reactivity of IgG glycans with Canavalia ensiformis agglutinin; PHA-L/Con A—a ratio between IgG N-glycan reactivity with PHA-L and IgG N-glycan reactivity with Con A; PHA-L— Phaseolus vulgaris leucoagglutinin, n—number of patients. For lectin specificity, see the Materials and Methods section. * IgG N-glycan reactivity with PHA-L was previously determined by us and presented as mean ± SD (standard deviation) in an article by Sołkiewicz et al. . Here, the obtained values are used for comparison with IgG N-glycan reactivities with Con A and the calculation of the PHA-L/Con A ratio. In the chart, the median is indicated as a line, colored boxes are 25–75% of results obtained, whiskers indicate min–max, and dots are outliers. A two-tailed p -value (probability value) of less than 0.05 was considered significant.
    Biotinylated Lectin Con A, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems biotinylated lotus tetragonolobus lectin
    a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus <t>tetragonolobus</t> <t>lectin,</t> E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).
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    Danaher Inc biotinylated lotus tetragonolobus lectin
    a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus <t>tetragonolobus</t> <t>lectin,</t> E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).
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    https://www.bioz.com/result/biotinylated lotus tetragonolobus lectin/product/Danaher Inc
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    a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus <t>tetragonolobus</t> <t>lectin,</t> E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).
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    https://www.bioz.com/result/biotinylated lotus tetragonolobus lectin/product/Vector Laboratories
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    Millipore biotinylated aleuria aurantia lectin aal
    a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus <t>tetragonolobus</t> <t>lectin,</t> E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).
    Biotinylated Aleuria Aurantia Lectin Aal, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories biotinylated sambucus nigra lectin
    (A-F) <t>Lectin</t> staining. <t>Biotinylated</t> <t>Sambucus</t> <t>Nigra</t> lectin (SNA) and Maackia Amurensis lectin II (MAL II) lectins were used to stain glycan expression in HEK293 SLC35A1-KO cells. NA-treated cells served as a positive control to show the removal of sialic acids. (A&B) Confocal microscopy . SNA (A) and MAL II (B) stained cells were incubated with DyLight 649-conjugated streptavidin for visualization at 100 × under a confocal microscope (Leica SP8 STED). (C&D) Flow cytometry. (C) SNA and (D) MAL II stained cells were incubated with FITC-conjugated streptavidin for flow cytometry. The histograms show the intensity of the FITC staining on the x-axis and the number of cells at each intensity level on the y-axis. The mean fluorescence intensity (MFI) values were calculated, normalized to the wild-type (WT) HEK293 cells as percentages (%), and are shown with a mean and SD from three replicates. P values were determined by using the Student’s t -test. (E-G) rAAV5 vector transduction, binding, and internalization in HEK293 cells. Relative percentages of vector binding (E), internalization (F), and transduction (G) to the Scramble cell group are calculated in rAAV-transduced SLC35A1-KO or NA-treated scramble HEK293 cells. The data shown were a mean and SD from three replicates. P values were determined by using one-way ANOVA for the comparison of the vector value in the KO or NA-treated cell group and the scramble cell group.
    Biotinylated Sambucus Nigra Lectin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated sambucus nigra lectin/product/Vector Laboratories
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    Image Search Results


    The values of relative reactivities of serum IgG N-glycans with * PHA-L, Con A, and PHA-L/Con A ratio. The results are expressed in absorbance units (AUs) as a median with an interquartile range (Q1–Q3). Serum IgG glycan reactivities with lectin were examined by lectin-ELISA and expressed in AU. Significant differences versus groups: 1 with advanced endometriosis (AE), 2 non-endometriosis (NE), and control—a group of healthy women. Con A—relative reactivity of IgG glycans with Canavalia ensiformis agglutinin; PHA-L/Con A—a ratio between IgG N-glycan reactivity with PHA-L and IgG N-glycan reactivity with Con A; PHA-L— Phaseolus vulgaris leucoagglutinin, n—number of patients. For lectin specificity, see the Materials and Methods section. * IgG N-glycan reactivity with PHA-L was previously determined by us and presented as mean ± SD (standard deviation) in an article by Sołkiewicz et al. . Here, the obtained values are used for comparison with IgG N-glycan reactivities with Con A and the calculation of the PHA-L/Con A ratio. In the chart, the median is indicated as a line, colored boxes are 25–75% of results obtained, whiskers indicate min–max, and dots are outliers. A two-tailed p -value (probability value) of less than 0.05 was considered significant.

    Journal: Molecules

    Article Title: The Degree of Branching of Serum IgG N-glycans as a Marker of Advanced Endometriosis

    doi: 10.3390/molecules29215136

    Figure Lengend Snippet: The values of relative reactivities of serum IgG N-glycans with * PHA-L, Con A, and PHA-L/Con A ratio. The results are expressed in absorbance units (AUs) as a median with an interquartile range (Q1–Q3). Serum IgG glycan reactivities with lectin were examined by lectin-ELISA and expressed in AU. Significant differences versus groups: 1 with advanced endometriosis (AE), 2 non-endometriosis (NE), and control—a group of healthy women. Con A—relative reactivity of IgG glycans with Canavalia ensiformis agglutinin; PHA-L/Con A—a ratio between IgG N-glycan reactivity with PHA-L and IgG N-glycan reactivity with Con A; PHA-L— Phaseolus vulgaris leucoagglutinin, n—number of patients. For lectin specificity, see the Materials and Methods section. * IgG N-glycan reactivity with PHA-L was previously determined by us and presented as mean ± SD (standard deviation) in an article by Sołkiewicz et al. . Here, the obtained values are used for comparison with IgG N-glycan reactivities with Con A and the calculation of the PHA-L/Con A ratio. In the chart, the median is indicated as a line, colored boxes are 25–75% of results obtained, whiskers indicate min–max, and dots are outliers. A two-tailed p -value (probability value) of less than 0.05 was considered significant.

    Article Snippet: The degree of branching of the IgG N-glycans was determined by a modified lectin-based ELISA with biotinylated lectin Con A (Vector Laboratories Inc., Burlingame, CA, USA) that recognizes α-linked mannose, specific for biantennary-type N-linked glycans.

    Techniques: Enzyme-linked Immunosorbent Assay, Control, Standard Deviation, Comparison, Two Tailed Test

    ROC curve analysis of IgG relative reactivity with  Con A,  and PHA-L/Con A ratio.

    Journal: Molecules

    Article Title: The Degree of Branching of Serum IgG N-glycans as a Marker of Advanced Endometriosis

    doi: 10.3390/molecules29215136

    Figure Lengend Snippet: ROC curve analysis of IgG relative reactivity with Con A, and PHA-L/Con A ratio.

    Article Snippet: The degree of branching of the IgG N-glycans was determined by a modified lectin-based ELISA with biotinylated lectin Con A (Vector Laboratories Inc., Burlingame, CA, USA) that recognizes α-linked mannose, specific for biantennary-type N-linked glycans.

    Techniques:

    a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus tetragonolobus lectin, E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).

    Journal: bioRxiv

    Article Title: Dynamic Kidney Organoid Microphysiological Analysis Platform

    doi: 10.1101/2024.10.27.620552

    Figure Lengend Snippet: a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus tetragonolobus lectin, E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).

    Article Snippet: The antibodies used in this study included anti-podocalyxin (AF1658, goat, R&D Systems, Inc), anti-E-Cadherin (ab11512, rat, Abcam), biotinylated Lotus tetragonolobus lectin (B-1325-2, Vector Laboratories), and anti-KIM-1 (MAB1750, mouse, R&D Systems, Inc), diluted at concentrations of 10µg·mL -1 , 5µg·mL -1 , 5µg·mL -1 , and 10µg·mL -1 , respectively.

    Techniques: Binding Assay, Construct, Confocal Microscopy, Expressing

    a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus tetragonolobus lectin, E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).

    Journal: bioRxiv

    Article Title: Dynamic Kidney Organoid Microphysiological Analysis Platform

    doi: 10.1101/2024.10.27.620552

    Figure Lengend Snippet: a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus tetragonolobus lectin, E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).

    Article Snippet: The antibodies used in this study included anti-podocalyxin (AF1658, goat, R&D Systems, Inc), anti-E-Cadherin (ab11512, rat, Abcam), biotinylated Lotus tetragonolobus lectin (B-1325-2, Vector Laboratories), and anti-KIM-1 (MAB1750, mouse, R&D Systems, Inc), diluted at concentrations of 10µg·mL -1 , 5µg·mL -1 , 5µg·mL -1 , and 10µg·mL -1 , respectively.

    Techniques: Binding Assay, Construct, Confocal Microscopy, Expressing

    a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus tetragonolobus lectin, E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).

    Journal: bioRxiv

    Article Title: Dynamic Kidney Organoid Microphysiological Analysis Platform

    doi: 10.1101/2024.10.27.620552

    Figure Lengend Snippet: a , The initial step of the MAP organogenesis, involving CHIR99021 priming of NPCs, emphasizes the crucial role of secondary signaling in addition to canonical Wnt/β-catenin signaling. The dynamic flow within MAP demonstrates that this signaling occurs in the context of secretion and receptor binding, functioning in an autocrine/paracrine manner. Computational and experimental results underscore the importance of secretion molecules, independent of Wnt/β-catenin easily governed by CHIR99201 perfusion, in facilitating the transition of NPCs into early epithelial nephron constructs. b , MAP’s precision in NPC differentiation signaling allows for the biochemical modulation of organoid development into either proximal-enriched or distal-enriched nephron development through the timely application of CHIR99021 and Y-27632. Optical monitoring within MAP exhibits the distinct developmental trajectories of these two types. Both patterns share a similar initial phase in NPC nephrogenesis, while tubules and stromal cells thrive in the later phases of the enhanced distal development. c , Structural analysis using confocal microscopy reveals the characteristic features of the proximal and the distal enrichment, especially significant distinction of distal tubules and stromal cells. The labels PODXL, LTL, ECAD, and DAPI correspond to podocalyxin, lotus tetragonolobus lectin, E-cadherin, and 4’,6-diamidino-2-phenylindole, respectively. d , The structural arrangement of MAP’s organoids with enhanced distal development reflects a sequential placement from PODXL-positive glomerulus-like structures, proximal tubules, and distal tubules with stroma, resembling early-stage kidney arrangement (Data are represented as the mean ± s.d. from 5 organoids in three independent experiments; data points excluded for visual clarity). e , Quantification of individual compartments indicates the prevalence of E-CAD positive distal tubules and the stromal population in enhanced distal development (EDD) compared to enhanced proximal development (EPD) (n EPD =10 and n EDD =5 from two and three independent experiments, respectively, and presented as the mean ± s.d). The normalized expression levels of the proximal tubule, distal tubule, and stroma showed a significance level in the t -test of p < 0.01 (***).

    Article Snippet: The antibodies used in this study included anti-podocalyxin (AF1658, goat, R&D Systems, Inc), anti-E-Cadherin (ab11512, rat, Abcam), biotinylated Lotus tetragonolobus lectin (B-1325-2, Vector Laboratories), and anti-KIM-1 (MAB1750, mouse, R&D Systems, Inc), diluted at concentrations of 10µg·mL -1 , 5µg·mL -1 , 5µg·mL -1 , and 10µg·mL -1 , respectively.

    Techniques: Binding Assay, Construct, Confocal Microscopy, Expressing

    (A-F) Lectin staining. Biotinylated Sambucus Nigra lectin (SNA) and Maackia Amurensis lectin II (MAL II) lectins were used to stain glycan expression in HEK293 SLC35A1-KO cells. NA-treated cells served as a positive control to show the removal of sialic acids. (A&B) Confocal microscopy . SNA (A) and MAL II (B) stained cells were incubated with DyLight 649-conjugated streptavidin for visualization at 100 × under a confocal microscope (Leica SP8 STED). (C&D) Flow cytometry. (C) SNA and (D) MAL II stained cells were incubated with FITC-conjugated streptavidin for flow cytometry. The histograms show the intensity of the FITC staining on the x-axis and the number of cells at each intensity level on the y-axis. The mean fluorescence intensity (MFI) values were calculated, normalized to the wild-type (WT) HEK293 cells as percentages (%), and are shown with a mean and SD from three replicates. P values were determined by using the Student’s t -test. (E-G) rAAV5 vector transduction, binding, and internalization in HEK293 cells. Relative percentages of vector binding (E), internalization (F), and transduction (G) to the Scramble cell group are calculated in rAAV-transduced SLC35A1-KO or NA-treated scramble HEK293 cells. The data shown were a mean and SD from three replicates. P values were determined by using one-way ANOVA for the comparison of the vector value in the KO or NA-treated cell group and the scramble cell group.

    Journal: bioRxiv

    Article Title: Identification of SLC35A1 as an essential host factor for the transduction of multi-serotype recombinant adeno-associated virus (AAV) vectors

    doi: 10.1101/2024.10.16.618764

    Figure Lengend Snippet: (A-F) Lectin staining. Biotinylated Sambucus Nigra lectin (SNA) and Maackia Amurensis lectin II (MAL II) lectins were used to stain glycan expression in HEK293 SLC35A1-KO cells. NA-treated cells served as a positive control to show the removal of sialic acids. (A&B) Confocal microscopy . SNA (A) and MAL II (B) stained cells were incubated with DyLight 649-conjugated streptavidin for visualization at 100 × under a confocal microscope (Leica SP8 STED). (C&D) Flow cytometry. (C) SNA and (D) MAL II stained cells were incubated with FITC-conjugated streptavidin for flow cytometry. The histograms show the intensity of the FITC staining on the x-axis and the number of cells at each intensity level on the y-axis. The mean fluorescence intensity (MFI) values were calculated, normalized to the wild-type (WT) HEK293 cells as percentages (%), and are shown with a mean and SD from three replicates. P values were determined by using the Student’s t -test. (E-G) rAAV5 vector transduction, binding, and internalization in HEK293 cells. Relative percentages of vector binding (E), internalization (F), and transduction (G) to the Scramble cell group are calculated in rAAV-transduced SLC35A1-KO or NA-treated scramble HEK293 cells. The data shown were a mean and SD from three replicates. P values were determined by using one-way ANOVA for the comparison of the vector value in the KO or NA-treated cell group and the scramble cell group.

    Article Snippet: Biotinylated Sambucus nigra lectin (SNA)(#B-1305), biotinylated Maackia amurensis lectin II (MAL II) (#B-1265), and fluorescein-conjugated Erythrina cristagalli lectin (ECL) (#FL-1141) were purchased from Vector Laboratories (Newark, CA).

    Techniques: Staining, Expressing, Positive Control, Confocal Microscopy, Incubation, Microscopy, Flow Cytometry, Fluorescence, Plasmid Preparation, Transduction, Binding Assay, Comparison

    (A&B) Confocal microscopy . SNA (A) and MALII (B) lectins were used to stain glycan expression in HAE-ALI SLC35A1-KO cultures. NA-treated cultures served as a positive control to show the removal of sialic acids. DyLight 649-conjugated streptavidin was used to visualize the staining under a confocal microscope at × 60 (CSU-W1 SoRa). (C&D) Flow cytometry of lection-stained cells dissociated from ALI cultures. (C) biotinylated SNA and (D) MAL II lectins were used to stain the cell surface, followed by FITC-conjugated streptavidin for detection. The histograms show the intensity of the FITC staining on the x-axis and the number of cells at each intensity level on the y-axis. The mean fluorescence intensity (MFI) values were calculated, normalized to the WT HEK293 cells. And the percentages (%) are shown with a mean and SD from three replicates. P values as indicated were determined by using the Student’s t -test.

    Journal: bioRxiv

    Article Title: Identification of SLC35A1 as an essential host factor for the transduction of multi-serotype recombinant adeno-associated virus (AAV) vectors

    doi: 10.1101/2024.10.16.618764

    Figure Lengend Snippet: (A&B) Confocal microscopy . SNA (A) and MALII (B) lectins were used to stain glycan expression in HAE-ALI SLC35A1-KO cultures. NA-treated cultures served as a positive control to show the removal of sialic acids. DyLight 649-conjugated streptavidin was used to visualize the staining under a confocal microscope at × 60 (CSU-W1 SoRa). (C&D) Flow cytometry of lection-stained cells dissociated from ALI cultures. (C) biotinylated SNA and (D) MAL II lectins were used to stain the cell surface, followed by FITC-conjugated streptavidin for detection. The histograms show the intensity of the FITC staining on the x-axis and the number of cells at each intensity level on the y-axis. The mean fluorescence intensity (MFI) values were calculated, normalized to the WT HEK293 cells. And the percentages (%) are shown with a mean and SD from three replicates. P values as indicated were determined by using the Student’s t -test.

    Article Snippet: Biotinylated Sambucus nigra lectin (SNA)(#B-1305), biotinylated Maackia amurensis lectin II (MAL II) (#B-1265), and fluorescein-conjugated Erythrina cristagalli lectin (ECL) (#FL-1141) were purchased from Vector Laboratories (Newark, CA).

    Techniques: Confocal Microscopy, Staining, Expressing, Positive Control, Microscopy, Flow Cytometry, Fluorescence

    (A) Luciferase activities. HEK293 Scramble and HEK293 SLC35A1-KO cells were respectively transduced with various serotypes of rAAV vectors as indicated at an MOI of 20,000 DRP/cell. At 3 dpt, the luciferase activities were measured and normalized to Scramble cells (set as 1). The fold changes of luciferase activities in SLC35A1-KO vs Scramble are shown with means and an SD from at least three replicates. (B&C) Lectin staining. HEK293 Scramble and HEK293 SLC35A1-KO cells were respectively stained with Erythrina cristagalli lectin (ECL) for analyses by confocal microscopy (B) and by flow cytometry (C). The mean fluorescence intensity (MFI) values were calculated and normalized to the Scramble cells as percentages (%), which are shown with a mean and SD from three replicates, and were analyzed by the Student’s t -test. (D-F) rAAV binding, internalization, transduction. HEK293 Scramble and HEK293 SLC35A1-KO cells were respectively transduced with four selected representative vectors, rAAV5, rAAV2, rAAV6, and rAAV9, in parallel. Vector binding (D), Internalization (E), and transduction (F) are assessed and relative fold changes in SLC35A1-KO vs Scramble as percentages (%) are shown with a mean and SD from at least three replicates. (G) Nuclear import assays . HEK293 Scramble and HEK293 SLC35A1-KO cells were respectively transduced with four selected representative vectors, rAAV5, rAAV2, rAAV6, and rAAV9, in parallel. At 12 hpt, the cytoplasm and nucleus were fractionated, and the percentage of vector genome copies in the cytoplasm and nucleus fractions were quantified. The data shown are means with an SD from at least three replicates. P value was determined by using the Student t -test for the comparison of the vector genome copies in the nucleus between the KO cell group and the Scramble cell group.

    Journal: bioRxiv

    Article Title: Identification of SLC35A1 as an essential host factor for the transduction of multi-serotype recombinant adeno-associated virus (AAV) vectors

    doi: 10.1101/2024.10.16.618764

    Figure Lengend Snippet: (A) Luciferase activities. HEK293 Scramble and HEK293 SLC35A1-KO cells were respectively transduced with various serotypes of rAAV vectors as indicated at an MOI of 20,000 DRP/cell. At 3 dpt, the luciferase activities were measured and normalized to Scramble cells (set as 1). The fold changes of luciferase activities in SLC35A1-KO vs Scramble are shown with means and an SD from at least three replicates. (B&C) Lectin staining. HEK293 Scramble and HEK293 SLC35A1-KO cells were respectively stained with Erythrina cristagalli lectin (ECL) for analyses by confocal microscopy (B) and by flow cytometry (C). The mean fluorescence intensity (MFI) values were calculated and normalized to the Scramble cells as percentages (%), which are shown with a mean and SD from three replicates, and were analyzed by the Student’s t -test. (D-F) rAAV binding, internalization, transduction. HEK293 Scramble and HEK293 SLC35A1-KO cells were respectively transduced with four selected representative vectors, rAAV5, rAAV2, rAAV6, and rAAV9, in parallel. Vector binding (D), Internalization (E), and transduction (F) are assessed and relative fold changes in SLC35A1-KO vs Scramble as percentages (%) are shown with a mean and SD from at least three replicates. (G) Nuclear import assays . HEK293 Scramble and HEK293 SLC35A1-KO cells were respectively transduced with four selected representative vectors, rAAV5, rAAV2, rAAV6, and rAAV9, in parallel. At 12 hpt, the cytoplasm and nucleus were fractionated, and the percentage of vector genome copies in the cytoplasm and nucleus fractions were quantified. The data shown are means with an SD from at least three replicates. P value was determined by using the Student t -test for the comparison of the vector genome copies in the nucleus between the KO cell group and the Scramble cell group.

    Article Snippet: Biotinylated Sambucus nigra lectin (SNA)(#B-1305), biotinylated Maackia amurensis lectin II (MAL II) (#B-1265), and fluorescein-conjugated Erythrina cristagalli lectin (ECL) (#FL-1141) were purchased from Vector Laboratories (Newark, CA).

    Techniques: Luciferase, Transduction, Staining, Confocal Microscopy, Flow Cytometry, Fluorescence, Binding Assay, Plasmid Preparation, Comparison

    HEK293 SLC35A1-KO cells were transduced with lentiviral vector that expressed SLC35A1 WT, T128A and ΔC Tail, as indicated, or untransduced (Mock), followed by selection of blasticidin (at 10 µg/ml) for a week. The blasticidin-resistant cells were transduced with rAAV5 at an MOI of 20,000. HEK293 Scramble cells were used as a control. (A) rAAV transduction efficiency. At 3 dpt, luciferase activities were measured and normalized to the Scramble (set as 1.0). Data shown are means with an SD from three replicates. P values were determined by using one-way ANOVA for the comparison of the fold changes in the SLC35A1 KO cell groups and the Scramble cell control. (B) rAAV genome distribution. After 12 hpt, nuclear and the cytoplasmic fractions of the rAAV5-transduced were fractionationed, and the vector genomes in each fraction were quantified by qPCR. The percentage of viral genome in each fraction shown are means with an SD of three replicates. P values were determined by using one-way ANOVA for the comparison of the vector genome copies in the nucleus between the SLC35A1 KO cell groups and the Scramble cell control. (C-F) Flow cytometry of lectin staining . The cells were stained with biotinylated SNA (C&D) or MALII (E&F) lectin and FITC-conjugated streptavidin, followed by flow cytometry. The mean fluorescence intensity (MFI) values were calculated, normalized to the WT HEK293 cells as percentages (%), and shown as means with an SD from at least three replicates. P values were determined by using one-way ANOVA for the comparison of the fold changes in the SLC35A1 KO cell groups and the Scramble cell control.

    Journal: bioRxiv

    Article Title: Identification of SLC35A1 as an essential host factor for the transduction of multi-serotype recombinant adeno-associated virus (AAV) vectors

    doi: 10.1101/2024.10.16.618764

    Figure Lengend Snippet: HEK293 SLC35A1-KO cells were transduced with lentiviral vector that expressed SLC35A1 WT, T128A and ΔC Tail, as indicated, or untransduced (Mock), followed by selection of blasticidin (at 10 µg/ml) for a week. The blasticidin-resistant cells were transduced with rAAV5 at an MOI of 20,000. HEK293 Scramble cells were used as a control. (A) rAAV transduction efficiency. At 3 dpt, luciferase activities were measured and normalized to the Scramble (set as 1.0). Data shown are means with an SD from three replicates. P values were determined by using one-way ANOVA for the comparison of the fold changes in the SLC35A1 KO cell groups and the Scramble cell control. (B) rAAV genome distribution. After 12 hpt, nuclear and the cytoplasmic fractions of the rAAV5-transduced were fractionationed, and the vector genomes in each fraction were quantified by qPCR. The percentage of viral genome in each fraction shown are means with an SD of three replicates. P values were determined by using one-way ANOVA for the comparison of the vector genome copies in the nucleus between the SLC35A1 KO cell groups and the Scramble cell control. (C-F) Flow cytometry of lectin staining . The cells were stained with biotinylated SNA (C&D) or MALII (E&F) lectin and FITC-conjugated streptavidin, followed by flow cytometry. The mean fluorescence intensity (MFI) values were calculated, normalized to the WT HEK293 cells as percentages (%), and shown as means with an SD from at least three replicates. P values were determined by using one-way ANOVA for the comparison of the fold changes in the SLC35A1 KO cell groups and the Scramble cell control.

    Article Snippet: Biotinylated Sambucus nigra lectin (SNA)(#B-1305), biotinylated Maackia amurensis lectin II (MAL II) (#B-1265), and fluorescein-conjugated Erythrina cristagalli lectin (ECL) (#FL-1141) were purchased from Vector Laboratories (Newark, CA).

    Techniques: Transduction, Plasmid Preparation, Selection, Control, Luciferase, Comparison, Flow Cytometry, Staining, Fluorescence