Review



dic8 pi 3 4 p2 p 3408  (Echelon Biosciences)


Bioz Verified Symbol Echelon Biosciences is a verified supplier
Bioz Manufacturer Symbol Echelon Biosciences manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Echelon Biosciences dic8 pi 3 4 p2 p 3408
    Dic8 Pi 3 4 P2 P 3408, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dic8 pi 3 4 p2 p 3408/product/Echelon Biosciences
    Average 94 stars, based on 1 article reviews
    dic8 pi 3 4 p2 p 3408 - by Bioz Stars, 2025-02
    94/100 stars

    Images



    Similar Products

    94
    Echelon Biosciences dic8 pi 3 4 p2 p 3408
    Dic8 Pi 3 4 P2 P 3408, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dic8 pi 3 4 p2 p 3408/product/Echelon Biosciences
    Average 94 stars, based on 1 article reviews
    dic8 pi 3 4 p2 p 3408 - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    94
    Echelon Biosciences dic8 pi 3 4 p 2 p 3408
    ( A ) The intensity of immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( B , C ) 0.05 μM His 6 -tagged PIPKIα ( B ) and IPMK ( C ) were incubated with 0.2 μM <t>diC8</t> PI(4)P or diC8 PI(4,5)P 2 , respectively, in the absence or presence of various concentrations of GST-YAP (0.001, 0.01, 0.1, 1.0, and 10.0 μM). The activities of the kinases were measured using the ADP-Glo assay (Promega). The graphs show the mean±s.d. of n = 3 independent experiments. YAP did not alter the activity of either kinase. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( D ) Summary of GST alone or GST-YAP bindings to PI, PI(4,5)P 2 , or PI(3,4,5)P 3 measured by MST. Raw data are available in Appendix Fig. S . ( E ) A schematic representation of the molecular structure Ac 3 2API and how it can become metabolically incorporated into phosphoinositides after removal of acetyl groups by esterase to produce azido- myo -inositol probe 2API. ( F ) The intensity of the immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( G ) Starved MDA-MB-231 cells were fed with Ac 3 2API for 24 h in the presence of 10% dialyzed serum. Cells were lysed and azide-tagged molecules were conjugated to biotin–alkyne through a click reaction. Endogenous c-Myc was immunoprecipitated and the associated complexes were analyzed by immunoblotting. Representative immunoblot images of n = 2 independent experiments are shown. ( H ) The clicked lysates were analyzed by anti-PI(4,5)P 2 or PI(3,4,5)P 3 antibodies. Biotinylated 2API was resolved by streptavidin. Many immunoblot bands overlapped with streptavidin signals. Representative immunoblot images of n = 3 independent experiments are shown. ( I , J ) Starved MDA-MB-231 cells were stimulated with 10% serum for 1 h. The images are z-stacks of PI(4,5)P 2 -YAP PLA ( E ) and PI(3,4,5)P 3 -YAP PLA ( F ) taken using a confocal microscope with each frame differing by 0.2 μm. DAPI was used to stain the nucleus. Representative images of n = 3 independent experiments are shown. Scale bar, 10 μm. ( K , L ) MDA-MB-231 cells grown in 10% serum were stimulated with 5 μM LPA for 90 min. Cells were fixed and the association of YAP with PI(4,5)P 2 ( G ) or PI(3,4,5)P 3 ( H ) was visualized by PLA. The images were obtained by widefield epifluorescence microscopy. The number of PLA puncta was counted from at least 10 cells and the graph shows the mean±s.d. of n = 3 independent experiments. DAPI staining was used to distinguish the nucleus from the cytoplasm. Treating the cells with LPA significantly increased the number of nuclear puncta. Scale bar, 10 μm. * P < 0.05; ** P < 0.01, and n.s; not significant in Student’s t test.
    Dic8 Pi 3 4 P 2 P 3408, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dic8 pi 3 4 p 2 p 3408/product/Echelon Biosciences
    Average 94 stars, based on 1 article reviews
    dic8 pi 3 4 p 2 p 3408 - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    94
    Echelon Biosciences 3 4 bisphosphate dic8
    ( A ) The intensity of immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( B , C ) 0.05 μM His 6 -tagged PIPKIα ( B ) and IPMK ( C ) were incubated with 0.2 μM <t>diC8</t> PI(4)P or diC8 PI(4,5)P 2 , respectively, in the absence or presence of various concentrations of GST-YAP (0.001, 0.01, 0.1, 1.0, and 10.0 μM). The activities of the kinases were measured using the ADP-Glo assay (Promega). The graphs show the mean±s.d. of n = 3 independent experiments. YAP did not alter the activity of either kinase. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( D ) Summary of GST alone or GST-YAP bindings to PI, PI(4,5)P 2 , or PI(3,4,5)P 3 measured by MST. Raw data are available in Appendix Fig. S . ( E ) A schematic representation of the molecular structure Ac 3 2API and how it can become metabolically incorporated into phosphoinositides after removal of acetyl groups by esterase to produce azido- myo -inositol probe 2API. ( F ) The intensity of the immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( G ) Starved MDA-MB-231 cells were fed with Ac 3 2API for 24 h in the presence of 10% dialyzed serum. Cells were lysed and azide-tagged molecules were conjugated to biotin–alkyne through a click reaction. Endogenous c-Myc was immunoprecipitated and the associated complexes were analyzed by immunoblotting. Representative immunoblot images of n = 2 independent experiments are shown. ( H ) The clicked lysates were analyzed by anti-PI(4,5)P 2 or PI(3,4,5)P 3 antibodies. Biotinylated 2API was resolved by streptavidin. Many immunoblot bands overlapped with streptavidin signals. Representative immunoblot images of n = 3 independent experiments are shown. ( I , J ) Starved MDA-MB-231 cells were stimulated with 10% serum for 1 h. The images are z-stacks of PI(4,5)P 2 -YAP PLA ( E ) and PI(3,4,5)P 3 -YAP PLA ( F ) taken using a confocal microscope with each frame differing by 0.2 μm. DAPI was used to stain the nucleus. Representative images of n = 3 independent experiments are shown. Scale bar, 10 μm. ( K , L ) MDA-MB-231 cells grown in 10% serum were stimulated with 5 μM LPA for 90 min. Cells were fixed and the association of YAP with PI(4,5)P 2 ( G ) or PI(3,4,5)P 3 ( H ) was visualized by PLA. The images were obtained by widefield epifluorescence microscopy. The number of PLA puncta was counted from at least 10 cells and the graph shows the mean±s.d. of n = 3 independent experiments. DAPI staining was used to distinguish the nucleus from the cytoplasm. Treating the cells with LPA significantly increased the number of nuclear puncta. Scale bar, 10 μm. * P < 0.05; ** P < 0.01, and n.s; not significant in Student’s t test.
    3 4 Bisphosphate Dic8, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/3 4 bisphosphate dic8/product/Echelon Biosciences
    Average 94 stars, based on 1 article reviews
    3 4 bisphosphate dic8 - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    94
    Echelon Biosciences pi(3,4)p2 dic8
    ( A ) The intensity of immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( B , C ) 0.05 μM His 6 -tagged PIPKIα ( B ) and IPMK ( C ) were incubated with 0.2 μM <t>diC8</t> PI(4)P or diC8 PI(4,5)P 2 , respectively, in the absence or presence of various concentrations of GST-YAP (0.001, 0.01, 0.1, 1.0, and 10.0 μM). The activities of the kinases were measured using the ADP-Glo assay (Promega). The graphs show the mean±s.d. of n = 3 independent experiments. YAP did not alter the activity of either kinase. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( D ) Summary of GST alone or GST-YAP bindings to PI, PI(4,5)P 2 , or PI(3,4,5)P 3 measured by MST. Raw data are available in Appendix Fig. S . ( E ) A schematic representation of the molecular structure Ac 3 2API and how it can become metabolically incorporated into phosphoinositides after removal of acetyl groups by esterase to produce azido- myo -inositol probe 2API. ( F ) The intensity of the immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( G ) Starved MDA-MB-231 cells were fed with Ac 3 2API for 24 h in the presence of 10% dialyzed serum. Cells were lysed and azide-tagged molecules were conjugated to biotin–alkyne through a click reaction. Endogenous c-Myc was immunoprecipitated and the associated complexes were analyzed by immunoblotting. Representative immunoblot images of n = 2 independent experiments are shown. ( H ) The clicked lysates were analyzed by anti-PI(4,5)P 2 or PI(3,4,5)P 3 antibodies. Biotinylated 2API was resolved by streptavidin. Many immunoblot bands overlapped with streptavidin signals. Representative immunoblot images of n = 3 independent experiments are shown. ( I , J ) Starved MDA-MB-231 cells were stimulated with 10% serum for 1 h. The images are z-stacks of PI(4,5)P 2 -YAP PLA ( E ) and PI(3,4,5)P 3 -YAP PLA ( F ) taken using a confocal microscope with each frame differing by 0.2 μm. DAPI was used to stain the nucleus. Representative images of n = 3 independent experiments are shown. Scale bar, 10 μm. ( K , L ) MDA-MB-231 cells grown in 10% serum were stimulated with 5 μM LPA for 90 min. Cells were fixed and the association of YAP with PI(4,5)P 2 ( G ) or PI(3,4,5)P 3 ( H ) was visualized by PLA. The images were obtained by widefield epifluorescence microscopy. The number of PLA puncta was counted from at least 10 cells and the graph shows the mean±s.d. of n = 3 independent experiments. DAPI staining was used to distinguish the nucleus from the cytoplasm. Treating the cells with LPA significantly increased the number of nuclear puncta. Scale bar, 10 μm. * P < 0.05; ** P < 0.01, and n.s; not significant in Student’s t test.
    Pi(3,4)P2 Dic8, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pi(3,4)p2 dic8/product/Echelon Biosciences
    Average 94 stars, based on 1 article reviews
    pi(3,4)p2 dic8 - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    86
    WILEY-VCH Verlag average 1 38 pd p 2 3264 10 2 3408 10
    ( A ) The intensity of immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( B , C ) 0.05 μM His 6 -tagged PIPKIα ( B ) and IPMK ( C ) were incubated with 0.2 μM <t>diC8</t> PI(4)P or diC8 PI(4,5)P 2 , respectively, in the absence or presence of various concentrations of GST-YAP (0.001, 0.01, 0.1, 1.0, and 10.0 μM). The activities of the kinases were measured using the ADP-Glo assay (Promega). The graphs show the mean±s.d. of n = 3 independent experiments. YAP did not alter the activity of either kinase. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( D ) Summary of GST alone or GST-YAP bindings to PI, PI(4,5)P 2 , or PI(3,4,5)P 3 measured by MST. Raw data are available in Appendix Fig. S . ( E ) A schematic representation of the molecular structure Ac 3 2API and how it can become metabolically incorporated into phosphoinositides after removal of acetyl groups by esterase to produce azido- myo -inositol probe 2API. ( F ) The intensity of the immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( G ) Starved MDA-MB-231 cells were fed with Ac 3 2API for 24 h in the presence of 10% dialyzed serum. Cells were lysed and azide-tagged molecules were conjugated to biotin–alkyne through a click reaction. Endogenous c-Myc was immunoprecipitated and the associated complexes were analyzed by immunoblotting. Representative immunoblot images of n = 2 independent experiments are shown. ( H ) The clicked lysates were analyzed by anti-PI(4,5)P 2 or PI(3,4,5)P 3 antibodies. Biotinylated 2API was resolved by streptavidin. Many immunoblot bands overlapped with streptavidin signals. Representative immunoblot images of n = 3 independent experiments are shown. ( I , J ) Starved MDA-MB-231 cells were stimulated with 10% serum for 1 h. The images are z-stacks of PI(4,5)P 2 -YAP PLA ( E ) and PI(3,4,5)P 3 -YAP PLA ( F ) taken using a confocal microscope with each frame differing by 0.2 μm. DAPI was used to stain the nucleus. Representative images of n = 3 independent experiments are shown. Scale bar, 10 μm. ( K , L ) MDA-MB-231 cells grown in 10% serum were stimulated with 5 μM LPA for 90 min. Cells were fixed and the association of YAP with PI(4,5)P 2 ( G ) or PI(3,4,5)P 3 ( H ) was visualized by PLA. The images were obtained by widefield epifluorescence microscopy. The number of PLA puncta was counted from at least 10 cells and the graph shows the mean±s.d. of n = 3 independent experiments. DAPI staining was used to distinguish the nucleus from the cytoplasm. Treating the cells with LPA significantly increased the number of nuclear puncta. Scale bar, 10 μm. * P < 0.05; ** P < 0.01, and n.s; not significant in Student’s t test.
    Average 1 38 Pd P 2 3264 10 2 3408 10, supplied by WILEY-VCH Verlag, 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/average 1 38 pd p 2 3264 10 2 3408 10/product/WILEY-VCH Verlag
    Average 86 stars, based on 1 article reviews
    average 1 38 pd p 2 3264 10 2 3408 10 - by Bioz Stars, 2025-02
    86/100 stars
      Buy from Supplier

    94
    Echelon Biosciences p 3408 pi 3 5 p dic8 echelon biosciences
    ( A ) The intensity of immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( B , C ) 0.05 μM His 6 -tagged PIPKIα ( B ) and IPMK ( C ) were incubated with 0.2 μM <t>diC8</t> PI(4)P or diC8 PI(4,5)P 2 , respectively, in the absence or presence of various concentrations of GST-YAP (0.001, 0.01, 0.1, 1.0, and 10.0 μM). The activities of the kinases were measured using the ADP-Glo assay (Promega). The graphs show the mean±s.d. of n = 3 independent experiments. YAP did not alter the activity of either kinase. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( D ) Summary of GST alone or GST-YAP bindings to PI, PI(4,5)P 2 , or PI(3,4,5)P 3 measured by MST. Raw data are available in Appendix Fig. S . ( E ) A schematic representation of the molecular structure Ac 3 2API and how it can become metabolically incorporated into phosphoinositides after removal of acetyl groups by esterase to produce azido- myo -inositol probe 2API. ( F ) The intensity of the immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( G ) Starved MDA-MB-231 cells were fed with Ac 3 2API for 24 h in the presence of 10% dialyzed serum. Cells were lysed and azide-tagged molecules were conjugated to biotin–alkyne through a click reaction. Endogenous c-Myc was immunoprecipitated and the associated complexes were analyzed by immunoblotting. Representative immunoblot images of n = 2 independent experiments are shown. ( H ) The clicked lysates were analyzed by anti-PI(4,5)P 2 or PI(3,4,5)P 3 antibodies. Biotinylated 2API was resolved by streptavidin. Many immunoblot bands overlapped with streptavidin signals. Representative immunoblot images of n = 3 independent experiments are shown. ( I , J ) Starved MDA-MB-231 cells were stimulated with 10% serum for 1 h. The images are z-stacks of PI(4,5)P 2 -YAP PLA ( E ) and PI(3,4,5)P 3 -YAP PLA ( F ) taken using a confocal microscope with each frame differing by 0.2 μm. DAPI was used to stain the nucleus. Representative images of n = 3 independent experiments are shown. Scale bar, 10 μm. ( K , L ) MDA-MB-231 cells grown in 10% serum were stimulated with 5 μM LPA for 90 min. Cells were fixed and the association of YAP with PI(4,5)P 2 ( G ) or PI(3,4,5)P 3 ( H ) was visualized by PLA. The images were obtained by widefield epifluorescence microscopy. The number of PLA puncta was counted from at least 10 cells and the graph shows the mean±s.d. of n = 3 independent experiments. DAPI staining was used to distinguish the nucleus from the cytoplasm. Treating the cells with LPA significantly increased the number of nuclear puncta. Scale bar, 10 μm. * P < 0.05; ** P < 0.01, and n.s; not significant in Student’s t test.
    P 3408 Pi 3 5 P Dic8 Echelon Biosciences, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p 3408 pi 3 5 p dic8 echelon biosciences/product/Echelon Biosciences
    Average 94 stars, based on 1 article reviews
    p 3408 pi 3 5 p dic8 echelon biosciences - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    94
    Echelon Biosciences p3408
    ( A ) The intensity of immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( B , C ) 0.05 μM His 6 -tagged PIPKIα ( B ) and IPMK ( C ) were incubated with 0.2 μM <t>diC8</t> PI(4)P or diC8 PI(4,5)P 2 , respectively, in the absence or presence of various concentrations of GST-YAP (0.001, 0.01, 0.1, 1.0, and 10.0 μM). The activities of the kinases were measured using the ADP-Glo assay (Promega). The graphs show the mean±s.d. of n = 3 independent experiments. YAP did not alter the activity of either kinase. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( D ) Summary of GST alone or GST-YAP bindings to PI, PI(4,5)P 2 , or PI(3,4,5)P 3 measured by MST. Raw data are available in Appendix Fig. S . ( E ) A schematic representation of the molecular structure Ac 3 2API and how it can become metabolically incorporated into phosphoinositides after removal of acetyl groups by esterase to produce azido- myo -inositol probe 2API. ( F ) The intensity of the immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( G ) Starved MDA-MB-231 cells were fed with Ac 3 2API for 24 h in the presence of 10% dialyzed serum. Cells were lysed and azide-tagged molecules were conjugated to biotin–alkyne through a click reaction. Endogenous c-Myc was immunoprecipitated and the associated complexes were analyzed by immunoblotting. Representative immunoblot images of n = 2 independent experiments are shown. ( H ) The clicked lysates were analyzed by anti-PI(4,5)P 2 or PI(3,4,5)P 3 antibodies. Biotinylated 2API was resolved by streptavidin. Many immunoblot bands overlapped with streptavidin signals. Representative immunoblot images of n = 3 independent experiments are shown. ( I , J ) Starved MDA-MB-231 cells were stimulated with 10% serum for 1 h. The images are z-stacks of PI(4,5)P 2 -YAP PLA ( E ) and PI(3,4,5)P 3 -YAP PLA ( F ) taken using a confocal microscope with each frame differing by 0.2 μm. DAPI was used to stain the nucleus. Representative images of n = 3 independent experiments are shown. Scale bar, 10 μm. ( K , L ) MDA-MB-231 cells grown in 10% serum were stimulated with 5 μM LPA for 90 min. Cells were fixed and the association of YAP with PI(4,5)P 2 ( G ) or PI(3,4,5)P 3 ( H ) was visualized by PLA. The images were obtained by widefield epifluorescence microscopy. The number of PLA puncta was counted from at least 10 cells and the graph shows the mean±s.d. of n = 3 independent experiments. DAPI staining was used to distinguish the nucleus from the cytoplasm. Treating the cells with LPA significantly increased the number of nuclear puncta. Scale bar, 10 μm. * P < 0.05; ** P < 0.01, and n.s; not significant in Student’s t test.
    P3408, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p3408/product/Echelon Biosciences
    Average 94 stars, based on 1 article reviews
    p3408 - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    94
    Echelon Biosciences pi 3 4 p 2 dic8
    a MCF-7 cells expressing GFP-INPP4B or GFP-vector were lysed, then subjected to whole cell proteomic analysis by LC-MS/MS. DAVID functional annotation bioinformatics microarray analysis was performed on upregulated proteins in GFP-INPP4B expressing cells versus GFP-vector cells ( n = 3 experiments). b MCF-7 cells expressing GFP-INPP4B or GFP-vector were serum-starved overnight then stimulated with EGF (100 ng/mL) for 5 min. Cells were lysed and subjected to immunoprecipitation using GFP-Trap beads. Bound proteins were identified by mass spectrometry using a cut-off of >2-fold change and p < 0.05 ( n = 2 experiments). c MCF-7 cells expressing GFP-INPP4B or GFP-vector were lysed and subjected to immunoprecipitation using GFP-Trap beads. Bound fractions and soluble lysates (5% of input) were subjected to immunoblotting to detect endogenous Rab7. d MCF-7 cells expressing GFP-INPP4B were serum starved overnight then stimulated with EGF (100 ng/mL) for 15 min. Cells were fixed and subjected to immuno-electron microscopy analysis using GFP (10 nm) and CD63 (15 nm) antibodies. Representative electron micrographs are shown. Arrows show GFP-INPP4B (white) and CD63 (black) localization. Yellow box indicates area where higher magnification micrograph was captured. e MCF-7 cells expressing HA-Rab7 WT , HA-Rab7 Q67L , or HA-Rab7 T22N were serum-starved overnight then stimulated with EGF (100 ng/mL) for 15 min. Cells were treated with saponin (0.02% w/v) to remove cytoplasmic proteins, then fixed and immunostained using INPP4B, CD63, and HA antibodies. f MCF-7 cells expressing GFP-INPP4B or GFP-vector were fixed and immunostained using purified recombinant GST-2xFYVE, DAPI, and phalloidin. Data represent the number of 2xFYVE+ puncta per cell ± SD ( n = 3 experiments, >50 cells per experiment). g MCF-7 cells expressing NT, INPP4B #1, or INPP4B #2 shRNA were fixed and immunostained with PI(3,4)P 2 antibodies, DAPI, and phalloidin. h , i MCF-7 cells expressing GFP-INPP4B or GFP-vector were fixed and immunostained using recombinant GST-2xFYVE and EEA1 ( h ) or CD63 ( i ) antibodies, and co-stained with DAPI. Data represent the percentage of 2xFYVE+ early endosomes ( h ) or late endosomes ( i ) ±SD ( n = 3 experiments, >30 cells per experiment). The inset panels at the lower right or under each image are higher power regions of the boxed areas. Scale bar is 200 nm ( d ), 10 µm ( e – i ). p values determined by a modified Fisher’s exact test are indicated in a , or by two-tailed unpaired t -test in b , f , i .
    Pi 3 4 P 2 Dic8, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pi 3 4 p 2 dic8/product/Echelon Biosciences
    Average 94 stars, based on 1 article reviews
    pi 3 4 p 2 dic8 - by Bioz Stars, 2025-02
    94/100 stars
      Buy from Supplier

    86
    Millipore p 5008 p 3408
    a MCF-7 cells expressing GFP-INPP4B or GFP-vector were lysed, then subjected to whole cell proteomic analysis by LC-MS/MS. DAVID functional annotation bioinformatics microarray analysis was performed on upregulated proteins in GFP-INPP4B expressing cells versus GFP-vector cells ( n = 3 experiments). b MCF-7 cells expressing GFP-INPP4B or GFP-vector were serum-starved overnight then stimulated with EGF (100 ng/mL) for 5 min. Cells were lysed and subjected to immunoprecipitation using GFP-Trap beads. Bound proteins were identified by mass spectrometry using a cut-off of >2-fold change and p < 0.05 ( n = 2 experiments). c MCF-7 cells expressing GFP-INPP4B or GFP-vector were lysed and subjected to immunoprecipitation using GFP-Trap beads. Bound fractions and soluble lysates (5% of input) were subjected to immunoblotting to detect endogenous Rab7. d MCF-7 cells expressing GFP-INPP4B were serum starved overnight then stimulated with EGF (100 ng/mL) for 15 min. Cells were fixed and subjected to immuno-electron microscopy analysis using GFP (10 nm) and CD63 (15 nm) antibodies. Representative electron micrographs are shown. Arrows show GFP-INPP4B (white) and CD63 (black) localization. Yellow box indicates area where higher magnification micrograph was captured. e MCF-7 cells expressing HA-Rab7 WT , HA-Rab7 Q67L , or HA-Rab7 T22N were serum-starved overnight then stimulated with EGF (100 ng/mL) for 15 min. Cells were treated with saponin (0.02% w/v) to remove cytoplasmic proteins, then fixed and immunostained using INPP4B, CD63, and HA antibodies. f MCF-7 cells expressing GFP-INPP4B or GFP-vector were fixed and immunostained using purified recombinant GST-2xFYVE, DAPI, and phalloidin. Data represent the number of 2xFYVE+ puncta per cell ± SD ( n = 3 experiments, >50 cells per experiment). g MCF-7 cells expressing NT, INPP4B #1, or INPP4B #2 shRNA were fixed and immunostained with PI(3,4)P 2 antibodies, DAPI, and phalloidin. h , i MCF-7 cells expressing GFP-INPP4B or GFP-vector were fixed and immunostained using recombinant GST-2xFYVE and EEA1 ( h ) or CD63 ( i ) antibodies, and co-stained with DAPI. Data represent the percentage of 2xFYVE+ early endosomes ( h ) or late endosomes ( i ) ±SD ( n = 3 experiments, >30 cells per experiment). The inset panels at the lower right or under each image are higher power regions of the boxed areas. Scale bar is 200 nm ( d ), 10 µm ( e – i ). p values determined by a modified Fisher’s exact test are indicated in a , or by two-tailed unpaired t -test in b , f , i .
    P 5008 P 3408, 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
    https://www.bioz.com/result/p 5008 p 3408/product/Millipore
    Average 86 stars, based on 1 article reviews
    p 5008 p 3408 - by Bioz Stars, 2025-02
    86/100 stars
      Buy from Supplier

    Image Search Results


    ( A ) The intensity of immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( B , C ) 0.05 μM His 6 -tagged PIPKIα ( B ) and IPMK ( C ) were incubated with 0.2 μM diC8 PI(4)P or diC8 PI(4,5)P 2 , respectively, in the absence or presence of various concentrations of GST-YAP (0.001, 0.01, 0.1, 1.0, and 10.0 μM). The activities of the kinases were measured using the ADP-Glo assay (Promega). The graphs show the mean±s.d. of n = 3 independent experiments. YAP did not alter the activity of either kinase. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( D ) Summary of GST alone or GST-YAP bindings to PI, PI(4,5)P 2 , or PI(3,4,5)P 3 measured by MST. Raw data are available in Appendix Fig. S . ( E ) A schematic representation of the molecular structure Ac 3 2API and how it can become metabolically incorporated into phosphoinositides after removal of acetyl groups by esterase to produce azido- myo -inositol probe 2API. ( F ) The intensity of the immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( G ) Starved MDA-MB-231 cells were fed with Ac 3 2API for 24 h in the presence of 10% dialyzed serum. Cells were lysed and azide-tagged molecules were conjugated to biotin–alkyne through a click reaction. Endogenous c-Myc was immunoprecipitated and the associated complexes were analyzed by immunoblotting. Representative immunoblot images of n = 2 independent experiments are shown. ( H ) The clicked lysates were analyzed by anti-PI(4,5)P 2 or PI(3,4,5)P 3 antibodies. Biotinylated 2API was resolved by streptavidin. Many immunoblot bands overlapped with streptavidin signals. Representative immunoblot images of n = 3 independent experiments are shown. ( I , J ) Starved MDA-MB-231 cells were stimulated with 10% serum for 1 h. The images are z-stacks of PI(4,5)P 2 -YAP PLA ( E ) and PI(3,4,5)P 3 -YAP PLA ( F ) taken using a confocal microscope with each frame differing by 0.2 μm. DAPI was used to stain the nucleus. Representative images of n = 3 independent experiments are shown. Scale bar, 10 μm. ( K , L ) MDA-MB-231 cells grown in 10% serum were stimulated with 5 μM LPA for 90 min. Cells were fixed and the association of YAP with PI(4,5)P 2 ( G ) or PI(3,4,5)P 3 ( H ) was visualized by PLA. The images were obtained by widefield epifluorescence microscopy. The number of PLA puncta was counted from at least 10 cells and the graph shows the mean±s.d. of n = 3 independent experiments. DAPI staining was used to distinguish the nucleus from the cytoplasm. Treating the cells with LPA significantly increased the number of nuclear puncta. Scale bar, 10 μm. * P < 0.05; ** P < 0.01, and n.s; not significant in Student’s t test.

    Journal: The EMBO Journal

    Article Title: Nuclear phosphoinositide signaling promotes YAP/TAZ-TEAD transcriptional activity in breast cancer

    doi: 10.1038/s44318-024-00085-6

    Figure Lengend Snippet: ( A ) The intensity of immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( B , C ) 0.05 μM His 6 -tagged PIPKIα ( B ) and IPMK ( C ) were incubated with 0.2 μM diC8 PI(4)P or diC8 PI(4,5)P 2 , respectively, in the absence or presence of various concentrations of GST-YAP (0.001, 0.01, 0.1, 1.0, and 10.0 μM). The activities of the kinases were measured using the ADP-Glo assay (Promega). The graphs show the mean±s.d. of n = 3 independent experiments. YAP did not alter the activity of either kinase. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( D ) Summary of GST alone or GST-YAP bindings to PI, PI(4,5)P 2 , or PI(3,4,5)P 3 measured by MST. Raw data are available in Appendix Fig. S . ( E ) A schematic representation of the molecular structure Ac 3 2API and how it can become metabolically incorporated into phosphoinositides after removal of acetyl groups by esterase to produce azido- myo -inositol probe 2API. ( F ) The intensity of the immunoblots of Fig. was quantified using ImageJ and the graph shows the mean ± s.d. of n = 3 independent experiments. * P < 0.05; ** P < 0.01, and n.s.; not significant in Student’s t test. ( G ) Starved MDA-MB-231 cells were fed with Ac 3 2API for 24 h in the presence of 10% dialyzed serum. Cells were lysed and azide-tagged molecules were conjugated to biotin–alkyne through a click reaction. Endogenous c-Myc was immunoprecipitated and the associated complexes were analyzed by immunoblotting. Representative immunoblot images of n = 2 independent experiments are shown. ( H ) The clicked lysates were analyzed by anti-PI(4,5)P 2 or PI(3,4,5)P 3 antibodies. Biotinylated 2API was resolved by streptavidin. Many immunoblot bands overlapped with streptavidin signals. Representative immunoblot images of n = 3 independent experiments are shown. ( I , J ) Starved MDA-MB-231 cells were stimulated with 10% serum for 1 h. The images are z-stacks of PI(4,5)P 2 -YAP PLA ( E ) and PI(3,4,5)P 3 -YAP PLA ( F ) taken using a confocal microscope with each frame differing by 0.2 μm. DAPI was used to stain the nucleus. Representative images of n = 3 independent experiments are shown. Scale bar, 10 μm. ( K , L ) MDA-MB-231 cells grown in 10% serum were stimulated with 5 μM LPA for 90 min. Cells were fixed and the association of YAP with PI(4,5)P 2 ( G ) or PI(3,4,5)P 3 ( H ) was visualized by PLA. The images were obtained by widefield epifluorescence microscopy. The number of PLA puncta was counted from at least 10 cells and the graph shows the mean±s.d. of n = 3 independent experiments. DAPI staining was used to distinguish the nucleus from the cytoplasm. Treating the cells with LPA significantly increased the number of nuclear puncta. Scale bar, 10 μm. * P < 0.05; ** P < 0.01, and n.s; not significant in Student’s t test.

    Article Snippet: Synthetic lipids diC8 PI (P-0008), diC8 PI(3)P (P-3008), diC8 PI(4)P (P-4008), diC8 PI(5)P (P-5008), diC8 PI(3,4)P 2 (P-3408), diC8 PI(3,5)P 2 (P-3508), diC8 PI(4,5)P 2 (P-4508), diC8 PI(3,4,5)P 3 (P-3908), diC16 PI(4,5)P 2 (P-4516), and diC16 PI(3,4,5)P 3 (P-3916) were purchased from Echelon Bioscience.

    Techniques: Western Blot, Incubation, Glo Assay, Activity Assay, Metabolic Labelling, Immunoprecipitation, Microscopy, Staining, Epifluorescence Microscopy

    a MCF-7 cells expressing GFP-INPP4B or GFP-vector were lysed, then subjected to whole cell proteomic analysis by LC-MS/MS. DAVID functional annotation bioinformatics microarray analysis was performed on upregulated proteins in GFP-INPP4B expressing cells versus GFP-vector cells ( n = 3 experiments). b MCF-7 cells expressing GFP-INPP4B or GFP-vector were serum-starved overnight then stimulated with EGF (100 ng/mL) for 5 min. Cells were lysed and subjected to immunoprecipitation using GFP-Trap beads. Bound proteins were identified by mass spectrometry using a cut-off of >2-fold change and p < 0.05 ( n = 2 experiments). c MCF-7 cells expressing GFP-INPP4B or GFP-vector were lysed and subjected to immunoprecipitation using GFP-Trap beads. Bound fractions and soluble lysates (5% of input) were subjected to immunoblotting to detect endogenous Rab7. d MCF-7 cells expressing GFP-INPP4B were serum starved overnight then stimulated with EGF (100 ng/mL) for 15 min. Cells were fixed and subjected to immuno-electron microscopy analysis using GFP (10 nm) and CD63 (15 nm) antibodies. Representative electron micrographs are shown. Arrows show GFP-INPP4B (white) and CD63 (black) localization. Yellow box indicates area where higher magnification micrograph was captured. e MCF-7 cells expressing HA-Rab7 WT , HA-Rab7 Q67L , or HA-Rab7 T22N were serum-starved overnight then stimulated with EGF (100 ng/mL) for 15 min. Cells were treated with saponin (0.02% w/v) to remove cytoplasmic proteins, then fixed and immunostained using INPP4B, CD63, and HA antibodies. f MCF-7 cells expressing GFP-INPP4B or GFP-vector were fixed and immunostained using purified recombinant GST-2xFYVE, DAPI, and phalloidin. Data represent the number of 2xFYVE+ puncta per cell ± SD ( n = 3 experiments, >50 cells per experiment). g MCF-7 cells expressing NT, INPP4B #1, or INPP4B #2 shRNA were fixed and immunostained with PI(3,4)P 2 antibodies, DAPI, and phalloidin. h , i MCF-7 cells expressing GFP-INPP4B or GFP-vector were fixed and immunostained using recombinant GST-2xFYVE and EEA1 ( h ) or CD63 ( i ) antibodies, and co-stained with DAPI. Data represent the percentage of 2xFYVE+ early endosomes ( h ) or late endosomes ( i ) ±SD ( n = 3 experiments, >30 cells per experiment). The inset panels at the lower right or under each image are higher power regions of the boxed areas. Scale bar is 200 nm ( d ), 10 µm ( e – i ). p values determined by a modified Fisher’s exact test are indicated in a , or by two-tailed unpaired t -test in b , f , i .

    Journal: Nature Communications

    Article Title: INPP4B promotes PI3Kα-dependent late endosome formation and Wnt/β-catenin signaling in breast cancer

    doi: 10.1038/s41467-021-23241-6

    Figure Lengend Snippet: a MCF-7 cells expressing GFP-INPP4B or GFP-vector were lysed, then subjected to whole cell proteomic analysis by LC-MS/MS. DAVID functional annotation bioinformatics microarray analysis was performed on upregulated proteins in GFP-INPP4B expressing cells versus GFP-vector cells ( n = 3 experiments). b MCF-7 cells expressing GFP-INPP4B or GFP-vector were serum-starved overnight then stimulated with EGF (100 ng/mL) for 5 min. Cells were lysed and subjected to immunoprecipitation using GFP-Trap beads. Bound proteins were identified by mass spectrometry using a cut-off of >2-fold change and p < 0.05 ( n = 2 experiments). c MCF-7 cells expressing GFP-INPP4B or GFP-vector were lysed and subjected to immunoprecipitation using GFP-Trap beads. Bound fractions and soluble lysates (5% of input) were subjected to immunoblotting to detect endogenous Rab7. d MCF-7 cells expressing GFP-INPP4B were serum starved overnight then stimulated with EGF (100 ng/mL) for 15 min. Cells were fixed and subjected to immuno-electron microscopy analysis using GFP (10 nm) and CD63 (15 nm) antibodies. Representative electron micrographs are shown. Arrows show GFP-INPP4B (white) and CD63 (black) localization. Yellow box indicates area where higher magnification micrograph was captured. e MCF-7 cells expressing HA-Rab7 WT , HA-Rab7 Q67L , or HA-Rab7 T22N were serum-starved overnight then stimulated with EGF (100 ng/mL) for 15 min. Cells were treated with saponin (0.02% w/v) to remove cytoplasmic proteins, then fixed and immunostained using INPP4B, CD63, and HA antibodies. f MCF-7 cells expressing GFP-INPP4B or GFP-vector were fixed and immunostained using purified recombinant GST-2xFYVE, DAPI, and phalloidin. Data represent the number of 2xFYVE+ puncta per cell ± SD ( n = 3 experiments, >50 cells per experiment). g MCF-7 cells expressing NT, INPP4B #1, or INPP4B #2 shRNA were fixed and immunostained with PI(3,4)P 2 antibodies, DAPI, and phalloidin. h , i MCF-7 cells expressing GFP-INPP4B or GFP-vector were fixed and immunostained using recombinant GST-2xFYVE and EEA1 ( h ) or CD63 ( i ) antibodies, and co-stained with DAPI. Data represent the percentage of 2xFYVE+ early endosomes ( h ) or late endosomes ( i ) ±SD ( n = 3 experiments, >30 cells per experiment). The inset panels at the lower right or under each image are higher power regions of the boxed areas. Scale bar is 200 nm ( d ), 10 µm ( e – i ). p values determined by a modified Fisher’s exact test are indicated in a , or by two-tailed unpaired t -test in b , f , i .

    Article Snippet: Samples were placed on ice for 2 min. About 50 µM PI(3,4)P 2 diC8 (Echelon, Cat # P-3408) was added and reactions were incubated at 37 °C for 20 min.

    Techniques: Expressing, Plasmid Preparation, Liquid Chromatography with Mass Spectroscopy, Functional Assay, Microarray, Immunoprecipitation, Mass Spectrometry, Western Blot, Immuno-Electron Microscopy, Purification, Recombinant, shRNA, Staining, Modification, Two Tailed Test

    a Expression of a panel of 17 Wnt pathway genes was assessed in 1469 ER + breast cancers from the METABRIC cohort, and stratified by PIK3CA -mutation status. The center line indicates the median, the lower bound of the box indicates the 25th percentile, the upper bound of the box represent the 75th percentile, the lower whisker extends from the 25th percentile to the 5th percentile, and the upper whisker extends from the 75th percentile to the 95th percentile. b – d MCF-7 cells expressing GFP-INPP4B or GFP-vector were treated with BKM120 (0.5 or 1 µM) or DMSO as a vehicle control for 48 h, then RNA was extracted two-step quantitative RT-PCR was performed using primers for AXIN2 ( b ), MYCN ( c ), or LEF1 ( d ). Expression was normalized to RRN18S . Expression was determined using the ΔΔCt method and expressed relative to vehicle-treated GFP-vector control cells (±SD), which were assigned an arbitrary value of 1 ( n = 3 experiments). e Model of PI3Kα and Wnt/β crosstalk via INPP4B-mediated late endosome formation. In PIK3CA -mutant ER + breast cancers, INPP4B is upregulated and localizes to the late endosome via its interaction with Rab7, which enhances its PI(3,4)P 2 4-phosphatase activity. INPP4B converts PI(3,4)P 2 to PI(3)P downstream of PI3Kα, which promotes Hrs-mediated late endosome formation, and increases the lysosomal degradation of GSK3β bound to the β-catenin destruction complex, promoting activation of Wnt/β-catenin signaling. p values determined by unpaired two-tailed Mann–Whitney test in a , or by one-way ANOVA with Tukey post hoc test in b , c , d .

    Journal: Nature Communications

    Article Title: INPP4B promotes PI3Kα-dependent late endosome formation and Wnt/β-catenin signaling in breast cancer

    doi: 10.1038/s41467-021-23241-6

    Figure Lengend Snippet: a Expression of a panel of 17 Wnt pathway genes was assessed in 1469 ER + breast cancers from the METABRIC cohort, and stratified by PIK3CA -mutation status. The center line indicates the median, the lower bound of the box indicates the 25th percentile, the upper bound of the box represent the 75th percentile, the lower whisker extends from the 25th percentile to the 5th percentile, and the upper whisker extends from the 75th percentile to the 95th percentile. b – d MCF-7 cells expressing GFP-INPP4B or GFP-vector were treated with BKM120 (0.5 or 1 µM) or DMSO as a vehicle control for 48 h, then RNA was extracted two-step quantitative RT-PCR was performed using primers for AXIN2 ( b ), MYCN ( c ), or LEF1 ( d ). Expression was normalized to RRN18S . Expression was determined using the ΔΔCt method and expressed relative to vehicle-treated GFP-vector control cells (±SD), which were assigned an arbitrary value of 1 ( n = 3 experiments). e Model of PI3Kα and Wnt/β crosstalk via INPP4B-mediated late endosome formation. In PIK3CA -mutant ER + breast cancers, INPP4B is upregulated and localizes to the late endosome via its interaction with Rab7, which enhances its PI(3,4)P 2 4-phosphatase activity. INPP4B converts PI(3,4)P 2 to PI(3)P downstream of PI3Kα, which promotes Hrs-mediated late endosome formation, and increases the lysosomal degradation of GSK3β bound to the β-catenin destruction complex, promoting activation of Wnt/β-catenin signaling. p values determined by unpaired two-tailed Mann–Whitney test in a , or by one-way ANOVA with Tukey post hoc test in b , c , d .

    Article Snippet: Samples were placed on ice for 2 min. About 50 µM PI(3,4)P 2 diC8 (Echelon, Cat # P-3408) was added and reactions were incubated at 37 °C for 20 min.

    Techniques: Expressing, Mutagenesis, Whisker Assay, Plasmid Preparation, Quantitative RT-PCR, Activity Assay, Activation Assay, Two Tailed Test, MANN-WHITNEY