human g6pd cdna  (OriGene)


Bioz Verified Symbol OriGene is a verified supplier
Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human g6pd cdna
    A <t>G6PD</t> protein expression was significantly decreased in diabetic kidney. The renal cortex from non-diabetic subjects (n=3) and diabetic patients (n=3) were examined by IHC staining for G6PD. Shown are average values with standard deviation (s.d.). **** denotes P < 0.0001 for DM versus NDM. B, C, D G6PD protein level was decreased in different diabetic rodents, including STZ diabetic rats (B), STZ diabetic mice (C) and Akita mice (D). The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were collected and Western blot was performed to examine the expression of G6PD protein. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM. E G6PD activity was decreased in the diabetic kidney. The renal cortex from non-diabetic (NDM) controls and STZ-induced diabetic mice (DM) were collected and G6PD activity was determined. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM.
    Human G6pd Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human g6pd cdna/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human g6pd cdna - by Bioz Stars, 2023-11
    88/100 stars

    Images

    1) Product Images from "High glucose-induced ubiquitylation of G6PD leads to the injury of podocyte"

    Article Title: High glucose-induced ubiquitylation of G6PD leads to the injury of podocyte

    Journal: bioRxiv

    doi: 10.1101/350694

    A G6PD protein expression was significantly decreased in diabetic kidney. The renal cortex from non-diabetic subjects (n=3) and diabetic patients (n=3) were examined by IHC staining for G6PD. Shown are average values with standard deviation (s.d.). **** denotes P < 0.0001 for DM versus NDM. B, C, D G6PD protein level was decreased in different diabetic rodents, including STZ diabetic rats (B), STZ diabetic mice (C) and Akita mice (D). The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were collected and Western blot was performed to examine the expression of G6PD protein. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM. E G6PD activity was decreased in the diabetic kidney. The renal cortex from non-diabetic (NDM) controls and STZ-induced diabetic mice (DM) were collected and G6PD activity was determined. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM.
    Figure Legend Snippet: A G6PD protein expression was significantly decreased in diabetic kidney. The renal cortex from non-diabetic subjects (n=3) and diabetic patients (n=3) were examined by IHC staining for G6PD. Shown are average values with standard deviation (s.d.). **** denotes P < 0.0001 for DM versus NDM. B, C, D G6PD protein level was decreased in different diabetic rodents, including STZ diabetic rats (B), STZ diabetic mice (C) and Akita mice (D). The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were collected and Western blot was performed to examine the expression of G6PD protein. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM. E G6PD activity was decreased in the diabetic kidney. The renal cortex from non-diabetic (NDM) controls and STZ-induced diabetic mice (DM) were collected and G6PD activity was determined. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM.

    Techniques Used: Expressing, Immunohistochemistry, Standard Deviation, Western Blot, Activity Assay

    A Podocyte number was decreased in diabetic kidney. The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were examined with immunofluorescence using anti-WT-1 antibody to label podocyte cells (green staining). The nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue staining). n=4 mice for each group. Magnification 40×. B G6PD protein level was decreased in podocytes of diabetic kidney. The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were examined with co-immunofluorescence using anti-WT-1 antibody (green staining) and anti-G6PD antibody (red staining). DAPI was used to label the nuclei (blue staining). Colocalization of the fluorochromes yielded a yellow color (see arrows). n=4 mice for each group. Magnification 40×.
    Figure Legend Snippet: A Podocyte number was decreased in diabetic kidney. The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were examined with immunofluorescence using anti-WT-1 antibody to label podocyte cells (green staining). The nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue staining). n=4 mice for each group. Magnification 40×. B G6PD protein level was decreased in podocytes of diabetic kidney. The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were examined with co-immunofluorescence using anti-WT-1 antibody (green staining) and anti-G6PD antibody (red staining). DAPI was used to label the nuclei (blue staining). Colocalization of the fluorochromes yielded a yellow color (see arrows). n=4 mice for each group. Magnification 40×.

    Techniques Used: Immunofluorescence, Staining

    A High glucose decreased G6PD protein level and increased podocytes apoptosis. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), or normal glucose supplemented with 19.4mM mannitol (NG+M) for 72 hours. Mannitol (M) was added as an osmotic control. The protein levels of G6PD and cleaved caspase-3 were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG or NG + M. B Palmitate decreased the expression of G6PD protein and increased the apoptosis of podocyte. Podocytes were treated with bovine serum albumin (BSA), 125μM palmitate (PA125), 250μM palmitate (PA250) or 500μM palmitate (PA500) for 24 hours. Protein levels of G6PD and cleaved caspase-3 were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA250 versus cells with BSA and ** denotes P < 0.01 for cells treated with PA500 versus cells with BSA.
    Figure Legend Snippet: A High glucose decreased G6PD protein level and increased podocytes apoptosis. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), or normal glucose supplemented with 19.4mM mannitol (NG+M) for 72 hours. Mannitol (M) was added as an osmotic control. The protein levels of G6PD and cleaved caspase-3 were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG or NG + M. B Palmitate decreased the expression of G6PD protein and increased the apoptosis of podocyte. Podocytes were treated with bovine serum albumin (BSA), 125μM palmitate (PA125), 250μM palmitate (PA250) or 500μM palmitate (PA500) for 24 hours. Protein levels of G6PD and cleaved caspase-3 were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA250 versus cells with BSA and ** denotes P < 0.01 for cells treated with PA500 versus cells with BSA.

    Techniques Used: Western Blot, Standard Deviation, Expressing

    A siRNA targeting to G6PD (siG6PD) significantly inhibited G6PD protein level. Podocytes were transfected with either siG6PD or scrambled siRNA for 48 hours and then G6PD protein expression was determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.0′5 for cells transfected with siG6PD versus cells with scramble. B Inhibition of G6PD led to the increased apoptosis of podocyte. Podocytes were transfected with either scrambled siRNA (scramble) or siG6PD and flow cytometry was used to detect podocytes apoptosis. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells transfected with siG6PD versus cells with scramble. C Deficiency of G6PD caused podocytes loss. The renal cortex from Hemi G6PD deficient mice aged 9wk, 23wk and 39wk and age-matched littermate wild type mice were examined with IHC staining for WT-1. n=6 mice for each group. Magnification 40×. ** denotes P < 0.01 and *** denotes P < 0.001 for Hemi G6PD deficient mice versus wild type mice. ### denotes P < 0.001 for 9wk Hemi G6PD deficient mice versus 39wk Hemi G6PD deficient mice. D Deficiency of G6PD caused increased podocyte apoptosis in vivo. The renal cortex from Hemizygous (Hemi) G6PD deficient mice (right panel) and age-matched littermate wild type mice (left panel) were examined with co-immunofluorescence using anti-WT-1 antibody to label podocyte cells (red staining) and TUNEL assay to label apoptotic cells (green staining). The nuclei were counterstained with DAPI (blue staining). Colocalization of the fluorochromes results in a yellow color (see arrows). n=6 mice for each group. Magnification 40×.
    Figure Legend Snippet: A siRNA targeting to G6PD (siG6PD) significantly inhibited G6PD protein level. Podocytes were transfected with either siG6PD or scrambled siRNA for 48 hours and then G6PD protein expression was determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.0′5 for cells transfected with siG6PD versus cells with scramble. B Inhibition of G6PD led to the increased apoptosis of podocyte. Podocytes were transfected with either scrambled siRNA (scramble) or siG6PD and flow cytometry was used to detect podocytes apoptosis. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells transfected with siG6PD versus cells with scramble. C Deficiency of G6PD caused podocytes loss. The renal cortex from Hemi G6PD deficient mice aged 9wk, 23wk and 39wk and age-matched littermate wild type mice were examined with IHC staining for WT-1. n=6 mice for each group. Magnification 40×. ** denotes P < 0.01 and *** denotes P < 0.001 for Hemi G6PD deficient mice versus wild type mice. ### denotes P < 0.001 for 9wk Hemi G6PD deficient mice versus 39wk Hemi G6PD deficient mice. D Deficiency of G6PD caused increased podocyte apoptosis in vivo. The renal cortex from Hemizygous (Hemi) G6PD deficient mice (right panel) and age-matched littermate wild type mice (left panel) were examined with co-immunofluorescence using anti-WT-1 antibody to label podocyte cells (red staining) and TUNEL assay to label apoptotic cells (green staining). The nuclei were counterstained with DAPI (blue staining). Colocalization of the fluorochromes results in a yellow color (see arrows). n=6 mice for each group. Magnification 40×.

    Techniques Used: Transfection, Expressing, Western Blot, Standard Deviation, Inhibition, Flow Cytometry, Immunohistochemistry, In Vivo, Immunofluorescence, Staining, TUNEL Assay

    A Overexpressing G6PD ameliorated podocytes apoptosis caused by high glucose. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), high glucose with infection of adenoviruses G6PD (HG+Ad-G6PD) and high glucose with infection of empty vector adenoviruses (HG+Ad-null). The levels of protein were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG. # denotes P < 0.05 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. B Elevation of G6PD rescued the apoptosis of podocyte induced by palmitate. Podocytes were treated with bovine serum albumin (BSA), 500μM palmitate (PA500), 500μM palmitate with infection of adenoviruses G6PD (PA500+Ad-G6PD) and 500μM palmitate with infection of empty vector adenoviruses (PA500+Ad-null). Protein expression were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA500 versus cells with BSA. # denotes P < 0.05 for cells treated with PA500+Ad-G6PD versus cells with PA500+Ad-null.
    Figure Legend Snippet: A Overexpressing G6PD ameliorated podocytes apoptosis caused by high glucose. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), high glucose with infection of adenoviruses G6PD (HG+Ad-G6PD) and high glucose with infection of empty vector adenoviruses (HG+Ad-null). The levels of protein were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG. # denotes P < 0.05 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. B Elevation of G6PD rescued the apoptosis of podocyte induced by palmitate. Podocytes were treated with bovine serum albumin (BSA), 500μM palmitate (PA500), 500μM palmitate with infection of adenoviruses G6PD (PA500+Ad-G6PD) and 500μM palmitate with infection of empty vector adenoviruses (PA500+Ad-null). Protein expression were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA500 versus cells with BSA. # denotes P < 0.05 for cells treated with PA500+Ad-G6PD versus cells with PA500+Ad-null.

    Techniques Used: Infection, Plasmid Preparation, Western Blot, Standard Deviation, Expressing

    A Overexpressing G6PD increased NADPH level in podocytes exposed to high glucose. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), high glucose with infection of adenoviruses G6PD (HG+Ad-G6PD) and high glucose with infection of empty vector adenoviruses (HG+Ad-null). NADPH was measured by a colorimetric method according to the manufacturer’s instructions. Shown are average values with standard deviation (s.d.) of triplicated experiments. ** denotes P < 0.01 for cells treated with HG versus cells with NG. # denotes P < 0.05 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. B the decreased GSH/GSSG in podocytes exposed to high glucose was ameliorated by overexpressing G6PD. Podocytes were treated as described in (A) and GSH/GSSG was measured by a spectrophotometric method following the manufacturer’s instructions. Shown are average values with standard deviation (s.d.) of triplicated experiments. ** denotes P < 0.01 for cells treated with HG versus cells with NG. ## denotes P < 0.01 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. C Elevation of G6PD protein expression reduced the accumulation of ROS in podocytes exposed to high glucose. Podocytes were treated as described in . ROS accumulation was measured with the cell-permeable, oxidation-sensitive dye CM-H 2 DCFDA. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG. ## denotes P < 0.01 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null.
    Figure Legend Snippet: A Overexpressing G6PD increased NADPH level in podocytes exposed to high glucose. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), high glucose with infection of adenoviruses G6PD (HG+Ad-G6PD) and high glucose with infection of empty vector adenoviruses (HG+Ad-null). NADPH was measured by a colorimetric method according to the manufacturer’s instructions. Shown are average values with standard deviation (s.d.) of triplicated experiments. ** denotes P < 0.01 for cells treated with HG versus cells with NG. # denotes P < 0.05 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. B the decreased GSH/GSSG in podocytes exposed to high glucose was ameliorated by overexpressing G6PD. Podocytes were treated as described in (A) and GSH/GSSG was measured by a spectrophotometric method following the manufacturer’s instructions. Shown are average values with standard deviation (s.d.) of triplicated experiments. ** denotes P < 0.01 for cells treated with HG versus cells with NG. ## denotes P < 0.01 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. C Elevation of G6PD protein expression reduced the accumulation of ROS in podocytes exposed to high glucose. Podocytes were treated as described in . ROS accumulation was measured with the cell-permeable, oxidation-sensitive dye CM-H 2 DCFDA. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG. ## denotes P < 0.01 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null.

    Techniques Used: Infection, Plasmid Preparation, Standard Deviation, Expressing

    A Palmitate decreased the level of G6PD mRNA. Podocytes were treated with BSA, 500μM palmitate (PA500) for 24 hours. G6PD mRNA level was measured by real-time PCR. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA500 versus cells with BSA. B High glucose had no effect on G6PD mRNA level. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), or normal glucose supplemented with mannitol (NG+M) for 72 hours. Real-time PCR was used to analyze G6PD mRNA abundance. Shown are average values with standard deviation (s.d.) of triplicated experiments. C The degradation of G6PD induced by high glucose was in a time-dependent manner. Podocytes were incubated with high glucose (HG, 25mM glucose) for the indicated time, and cell lysates were subjected to G6PD and β-actin immunoblotting. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG for 72 hours versus cells incubated in normal glucose. D The decreased G6PD protein level in high glucose was largely rescued by MG132. Podocytes were cultured with normal glucose (NG, 5.6mM glucose), normal glucose supplemented with 0.5μM MG132 (NG+MG132), high glucose (HG, 25mM glucose) or high glucose supplemented with 0.5μM MG132 (HG+MG132). Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with HG+MG132. E G6PD was ubiquitylated. HEK293T cells were transfected with indicated plasmids with or without 20μM MG132 for 12 hours. Cell lysates were subjected to immunoprecipitation (IP) with anti-Flag M2 beads. The precipitates were probed using His and Flag antibodies. Input cell lysates were subjected to Flag and GAPDH immunoblotting.
    Figure Legend Snippet: A Palmitate decreased the level of G6PD mRNA. Podocytes were treated with BSA, 500μM palmitate (PA500) for 24 hours. G6PD mRNA level was measured by real-time PCR. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA500 versus cells with BSA. B High glucose had no effect on G6PD mRNA level. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), or normal glucose supplemented with mannitol (NG+M) for 72 hours. Real-time PCR was used to analyze G6PD mRNA abundance. Shown are average values with standard deviation (s.d.) of triplicated experiments. C The degradation of G6PD induced by high glucose was in a time-dependent manner. Podocytes were incubated with high glucose (HG, 25mM glucose) for the indicated time, and cell lysates were subjected to G6PD and β-actin immunoblotting. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG for 72 hours versus cells incubated in normal glucose. D The decreased G6PD protein level in high glucose was largely rescued by MG132. Podocytes were cultured with normal glucose (NG, 5.6mM glucose), normal glucose supplemented with 0.5μM MG132 (NG+MG132), high glucose (HG, 25mM glucose) or high glucose supplemented with 0.5μM MG132 (HG+MG132). Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with HG+MG132. E G6PD was ubiquitylated. HEK293T cells were transfected with indicated plasmids with or without 20μM MG132 for 12 hours. Cell lysates were subjected to immunoprecipitation (IP) with anti-Flag M2 beads. The precipitates were probed using His and Flag antibodies. Input cell lysates were subjected to Flag and GAPDH immunoblotting.

    Techniques Used: Real-time Polymerase Chain Reaction, Standard Deviation, Incubation, Western Blot, Cell Culture, Transfection, Immunoprecipitation

    A Flag-G6PD plasmid was expressed in HEK293T cells. Flag-G6PD plasmid was transfected into HEK293T cells for 48 hours. Cells lysates were subjected to Flag and GAPDH immunoblotting. B His-Ub plasmid was verified in HEK293T cells. HEK293T cells were transfected with His-Ub plasmid for 48 hours. Protein level of His was analyzed by Western blot with anti-His antibody. C HA-VHL plasmid was expressed in HEK293T. HEK293T cells were transfected with HA-VHL plasmid for 48 hours and cells lysates were subjected to HA and GAPDH immunoblotting.
    Figure Legend Snippet: A Flag-G6PD plasmid was expressed in HEK293T cells. Flag-G6PD plasmid was transfected into HEK293T cells for 48 hours. Cells lysates were subjected to Flag and GAPDH immunoblotting. B His-Ub plasmid was verified in HEK293T cells. HEK293T cells were transfected with His-Ub plasmid for 48 hours. Protein level of His was analyzed by Western blot with anti-His antibody. C HA-VHL plasmid was expressed in HEK293T. HEK293T cells were transfected with HA-VHL plasmid for 48 hours and cells lysates were subjected to HA and GAPDH immunoblotting.

    Techniques Used: Plasmid Preparation, Transfection, Western Blot

    A Human G6PD interacted with the E3 ubiquitin ligase VHL in Y2H system. Using human G6PD as bait, VHL was identified as an interacting protein with G6PD in yeast. G6PD and VHL were co-transformed into yeast strain Mav203-activated expression of β-galactosidase. AD, Activation Domain; BD, Binding Domain; SD-2, deficient in Leucine and Tryptophan; SD-4, deficient in Leucine, Tryptophan, Histidine, and Uracil. B G6PD interacted with VHL. Co-immunoprecipitation assay shown that tagged G6PD and VHL formed a complex in HEK293T cells. HEK293T cells were transfected with HA-VHL expression plasmid in combination with or without Flag-G6PD expression plasmid. Flag-G6PD was immunoprecipitated with anti-Flag M2 beads. The precipitates were probed using HA and Flag antibodies. C VHL negatively regulated the expression of G6PD protein. HEK293T cells were co-transfected with Flag-G6PD expression plasmid and increasing amounts of HA-VHL expression plasmid. The levels of Flag and HA were determined by Western blot with indicated antibodies. D VHL reduced G6PD protein stability. HEK293T cells were transfected with Flag-G6PD expression plasmid with or without HA-VHL expression plasmid. After 36 hours, cells were treated with CHX (100μg/ml) for the indicated time. Cell lysates were subjected to Flag, GAPDH and HA immunoblotting. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells co-transfected with Flag-G6PD and HA-VHL (Flag-G6PD + HA-VHL) versus cells transfected with Flag-G6PD. E G6PD was efficiently ubiquitylated in the presence of VHL. HEK293T cells were transfected with different combinations of plasmids as indicated. G6PD was immunoprecipitated with anti-Flag M2 beads and immunoblotted with anti-His antibody to detect ubiquitylated G6PD. F Knockdown of endogenous VHL enhanced G6PD protein abundance. HEK293T cells were transfected with scrambled siRNA (scramble), siVHL#1 and siVHL#2. The levels of endogenous G6PD and VHL were analyzed by Western blot with indicated antibodies. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with siVHL#2 versus cells with scramble.
    Figure Legend Snippet: A Human G6PD interacted with the E3 ubiquitin ligase VHL in Y2H system. Using human G6PD as bait, VHL was identified as an interacting protein with G6PD in yeast. G6PD and VHL were co-transformed into yeast strain Mav203-activated expression of β-galactosidase. AD, Activation Domain; BD, Binding Domain; SD-2, deficient in Leucine and Tryptophan; SD-4, deficient in Leucine, Tryptophan, Histidine, and Uracil. B G6PD interacted with VHL. Co-immunoprecipitation assay shown that tagged G6PD and VHL formed a complex in HEK293T cells. HEK293T cells were transfected with HA-VHL expression plasmid in combination with or without Flag-G6PD expression plasmid. Flag-G6PD was immunoprecipitated with anti-Flag M2 beads. The precipitates were probed using HA and Flag antibodies. C VHL negatively regulated the expression of G6PD protein. HEK293T cells were co-transfected with Flag-G6PD expression plasmid and increasing amounts of HA-VHL expression plasmid. The levels of Flag and HA were determined by Western blot with indicated antibodies. D VHL reduced G6PD protein stability. HEK293T cells were transfected with Flag-G6PD expression plasmid with or without HA-VHL expression plasmid. After 36 hours, cells were treated with CHX (100μg/ml) for the indicated time. Cell lysates were subjected to Flag, GAPDH and HA immunoblotting. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells co-transfected with Flag-G6PD and HA-VHL (Flag-G6PD + HA-VHL) versus cells transfected with Flag-G6PD. E G6PD was efficiently ubiquitylated in the presence of VHL. HEK293T cells were transfected with different combinations of plasmids as indicated. G6PD was immunoprecipitated with anti-Flag M2 beads and immunoblotted with anti-His antibody to detect ubiquitylated G6PD. F Knockdown of endogenous VHL enhanced G6PD protein abundance. HEK293T cells were transfected with scrambled siRNA (scramble), siVHL#1 and siVHL#2. The levels of endogenous G6PD and VHL were analyzed by Western blot with indicated antibodies. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with siVHL#2 versus cells with scramble.

    Techniques Used: Transformation Assay, Expressing, Activation Assay, Binding Assay, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Standard Deviation

    A The map of the 5 lysine sites for VHL mediated ubiquitylation on G6PD. HEK293T cells were transfected with Flag-G6PD, His-Ub and HA-VHL plasmids, and then ubiquitylated G6PD were immunoprecipitated with anti-Flag M2 beads. MS spectra analysis identified 5 Lys residues (K 97 , K 366 , K 403 , K 407 and K 408 ) for VHL-mediated ubiquitylation in G6PD. B K 366 and K 403 were the major sites for VHL-mediated ubiquitylation on G6PD. HEK293T cells were transfected with Flag-tagged wild-type G6PD (WT) or its mutants bearing single Lys-to-Arg substitutions at the above 5 potential ubiquitylation sites. Cell lysates were subjected to immunoprecipitation (IP) with anti-Flag M2 beads. The precipitates were probed using His and Flag antibodies. C Mutants with both K 366 and K 403 sites (K 366 + 403R ) largely abolished the ubiquitylation of G6PD. HEK293T cells were transfected with Flag-tagged wild-type G6PD (WT) or G6PD mutants bearing single or two Lys-to-Arg substitutions at K 366 and K 403 sites.
    Figure Legend Snippet: A The map of the 5 lysine sites for VHL mediated ubiquitylation on G6PD. HEK293T cells were transfected with Flag-G6PD, His-Ub and HA-VHL plasmids, and then ubiquitylated G6PD were immunoprecipitated with anti-Flag M2 beads. MS spectra analysis identified 5 Lys residues (K 97 , K 366 , K 403 , K 407 and K 408 ) for VHL-mediated ubiquitylation in G6PD. B K 366 and K 403 were the major sites for VHL-mediated ubiquitylation on G6PD. HEK293T cells were transfected with Flag-tagged wild-type G6PD (WT) or its mutants bearing single Lys-to-Arg substitutions at the above 5 potential ubiquitylation sites. Cell lysates were subjected to immunoprecipitation (IP) with anti-Flag M2 beads. The precipitates were probed using His and Flag antibodies. C Mutants with both K 366 and K 403 sites (K 366 + 403R ) largely abolished the ubiquitylation of G6PD. HEK293T cells were transfected with Flag-tagged wild-type G6PD (WT) or G6PD mutants bearing single or two Lys-to-Arg substitutions at K 366 and K 403 sites.

    Techniques Used: Transfection, Immunoprecipitation

    human g6pd cdna  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human g6pd cdna
    A <t>G6PD</t> protein expression was significantly decreased in diabetic kidney. The renal cortex from non-diabetic subjects (n=3) and diabetic patients (n=3) were examined by IHC staining for G6PD. Shown are average values with standard deviation (s.d.). **** denotes P < 0.0001 for DM versus NDM. B, C, D G6PD protein level was decreased in different diabetic rodents, including STZ diabetic rats (B), STZ diabetic mice (C) and Akita mice (D). The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were collected and Western blot was performed to examine the expression of G6PD protein. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM. E G6PD activity was decreased in the diabetic kidney. The renal cortex from non-diabetic (NDM) controls and STZ-induced diabetic mice (DM) were collected and G6PD activity was determined. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM.
    Human G6pd Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human g6pd cdna/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human g6pd cdna - by Bioz Stars, 2023-11
    88/100 stars

    Images

    1) Product Images from "High glucose-induced ubiquitylation of G6PD leads to the injury of podocyte"

    Article Title: High glucose-induced ubiquitylation of G6PD leads to the injury of podocyte

    Journal: bioRxiv

    doi: 10.1101/350694

    A G6PD protein expression was significantly decreased in diabetic kidney. The renal cortex from non-diabetic subjects (n=3) and diabetic patients (n=3) were examined by IHC staining for G6PD. Shown are average values with standard deviation (s.d.). **** denotes P < 0.0001 for DM versus NDM. B, C, D G6PD protein level was decreased in different diabetic rodents, including STZ diabetic rats (B), STZ diabetic mice (C) and Akita mice (D). The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were collected and Western blot was performed to examine the expression of G6PD protein. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM. E G6PD activity was decreased in the diabetic kidney. The renal cortex from non-diabetic (NDM) controls and STZ-induced diabetic mice (DM) were collected and G6PD activity was determined. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM.
    Figure Legend Snippet: A G6PD protein expression was significantly decreased in diabetic kidney. The renal cortex from non-diabetic subjects (n=3) and diabetic patients (n=3) were examined by IHC staining for G6PD. Shown are average values with standard deviation (s.d.). **** denotes P < 0.0001 for DM versus NDM. B, C, D G6PD protein level was decreased in different diabetic rodents, including STZ diabetic rats (B), STZ diabetic mice (C) and Akita mice (D). The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were collected and Western blot was performed to examine the expression of G6PD protein. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM. E G6PD activity was decreased in the diabetic kidney. The renal cortex from non-diabetic (NDM) controls and STZ-induced diabetic mice (DM) were collected and G6PD activity was determined. Shown are average values with standard deviation (s.d.). n=5 mice for each group. * denotes P < 0.05 for DM versus NDM.

    Techniques Used: Expressing, Immunohistochemistry, Standard Deviation, Western Blot, Activity Assay

    A Podocyte number was decreased in diabetic kidney. The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were examined with immunofluorescence using anti-WT-1 antibody to label podocyte cells (green staining). The nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue staining). n=4 mice for each group. Magnification 40×. B G6PD protein level was decreased in podocytes of diabetic kidney. The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were examined with co-immunofluorescence using anti-WT-1 antibody (green staining) and anti-G6PD antibody (red staining). DAPI was used to label the nuclei (blue staining). Colocalization of the fluorochromes yielded a yellow color (see arrows). n=4 mice for each group. Magnification 40×.
    Figure Legend Snippet: A Podocyte number was decreased in diabetic kidney. The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were examined with immunofluorescence using anti-WT-1 antibody to label podocyte cells (green staining). The nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue staining). n=4 mice for each group. Magnification 40×. B G6PD protein level was decreased in podocytes of diabetic kidney. The renal cortex from non-diabetic (NDM) controls and diabetic rodents (DM) were examined with co-immunofluorescence using anti-WT-1 antibody (green staining) and anti-G6PD antibody (red staining). DAPI was used to label the nuclei (blue staining). Colocalization of the fluorochromes yielded a yellow color (see arrows). n=4 mice for each group. Magnification 40×.

    Techniques Used: Immunofluorescence, Staining

    A High glucose decreased G6PD protein level and increased podocytes apoptosis. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), or normal glucose supplemented with 19.4mM mannitol (NG+M) for 72 hours. Mannitol (M) was added as an osmotic control. The protein levels of G6PD and cleaved caspase-3 were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG or NG + M. B Palmitate decreased the expression of G6PD protein and increased the apoptosis of podocyte. Podocytes were treated with bovine serum albumin (BSA), 125μM palmitate (PA125), 250μM palmitate (PA250) or 500μM palmitate (PA500) for 24 hours. Protein levels of G6PD and cleaved caspase-3 were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA250 versus cells with BSA and ** denotes P < 0.01 for cells treated with PA500 versus cells with BSA.
    Figure Legend Snippet: A High glucose decreased G6PD protein level and increased podocytes apoptosis. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), or normal glucose supplemented with 19.4mM mannitol (NG+M) for 72 hours. Mannitol (M) was added as an osmotic control. The protein levels of G6PD and cleaved caspase-3 were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG or NG + M. B Palmitate decreased the expression of G6PD protein and increased the apoptosis of podocyte. Podocytes were treated with bovine serum albumin (BSA), 125μM palmitate (PA125), 250μM palmitate (PA250) or 500μM palmitate (PA500) for 24 hours. Protein levels of G6PD and cleaved caspase-3 were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA250 versus cells with BSA and ** denotes P < 0.01 for cells treated with PA500 versus cells with BSA.

    Techniques Used: Western Blot, Standard Deviation, Expressing

    A siRNA targeting to G6PD (siG6PD) significantly inhibited G6PD protein level. Podocytes were transfected with either siG6PD or scrambled siRNA for 48 hours and then G6PD protein expression was determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.0′5 for cells transfected with siG6PD versus cells with scramble. B Inhibition of G6PD led to the increased apoptosis of podocyte. Podocytes were transfected with either scrambled siRNA (scramble) or siG6PD and flow cytometry was used to detect podocytes apoptosis. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells transfected with siG6PD versus cells with scramble. C Deficiency of G6PD caused podocytes loss. The renal cortex from Hemi G6PD deficient mice aged 9wk, 23wk and 39wk and age-matched littermate wild type mice were examined with IHC staining for WT-1. n=6 mice for each group. Magnification 40×. ** denotes P < 0.01 and *** denotes P < 0.001 for Hemi G6PD deficient mice versus wild type mice. ### denotes P < 0.001 for 9wk Hemi G6PD deficient mice versus 39wk Hemi G6PD deficient mice. D Deficiency of G6PD caused increased podocyte apoptosis in vivo. The renal cortex from Hemizygous (Hemi) G6PD deficient mice (right panel) and age-matched littermate wild type mice (left panel) were examined with co-immunofluorescence using anti-WT-1 antibody to label podocyte cells (red staining) and TUNEL assay to label apoptotic cells (green staining). The nuclei were counterstained with DAPI (blue staining). Colocalization of the fluorochromes results in a yellow color (see arrows). n=6 mice for each group. Magnification 40×.
    Figure Legend Snippet: A siRNA targeting to G6PD (siG6PD) significantly inhibited G6PD protein level. Podocytes were transfected with either siG6PD or scrambled siRNA for 48 hours and then G6PD protein expression was determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.0′5 for cells transfected with siG6PD versus cells with scramble. B Inhibition of G6PD led to the increased apoptosis of podocyte. Podocytes were transfected with either scrambled siRNA (scramble) or siG6PD and flow cytometry was used to detect podocytes apoptosis. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells transfected with siG6PD versus cells with scramble. C Deficiency of G6PD caused podocytes loss. The renal cortex from Hemi G6PD deficient mice aged 9wk, 23wk and 39wk and age-matched littermate wild type mice were examined with IHC staining for WT-1. n=6 mice for each group. Magnification 40×. ** denotes P < 0.01 and *** denotes P < 0.001 for Hemi G6PD deficient mice versus wild type mice. ### denotes P < 0.001 for 9wk Hemi G6PD deficient mice versus 39wk Hemi G6PD deficient mice. D Deficiency of G6PD caused increased podocyte apoptosis in vivo. The renal cortex from Hemizygous (Hemi) G6PD deficient mice (right panel) and age-matched littermate wild type mice (left panel) were examined with co-immunofluorescence using anti-WT-1 antibody to label podocyte cells (red staining) and TUNEL assay to label apoptotic cells (green staining). The nuclei were counterstained with DAPI (blue staining). Colocalization of the fluorochromes results in a yellow color (see arrows). n=6 mice for each group. Magnification 40×.

    Techniques Used: Transfection, Expressing, Western Blot, Standard Deviation, Inhibition, Flow Cytometry, Immunohistochemistry, In Vivo, Immunofluorescence, Staining, TUNEL Assay

    A Overexpressing G6PD ameliorated podocytes apoptosis caused by high glucose. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), high glucose with infection of adenoviruses G6PD (HG+Ad-G6PD) and high glucose with infection of empty vector adenoviruses (HG+Ad-null). The levels of protein were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG. # denotes P < 0.05 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. B Elevation of G6PD rescued the apoptosis of podocyte induced by palmitate. Podocytes were treated with bovine serum albumin (BSA), 500μM palmitate (PA500), 500μM palmitate with infection of adenoviruses G6PD (PA500+Ad-G6PD) and 500μM palmitate with infection of empty vector adenoviruses (PA500+Ad-null). Protein expression were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA500 versus cells with BSA. # denotes P < 0.05 for cells treated with PA500+Ad-G6PD versus cells with PA500+Ad-null.
    Figure Legend Snippet: A Overexpressing G6PD ameliorated podocytes apoptosis caused by high glucose. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), high glucose with infection of adenoviruses G6PD (HG+Ad-G6PD) and high glucose with infection of empty vector adenoviruses (HG+Ad-null). The levels of protein were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG. # denotes P < 0.05 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. B Elevation of G6PD rescued the apoptosis of podocyte induced by palmitate. Podocytes were treated with bovine serum albumin (BSA), 500μM palmitate (PA500), 500μM palmitate with infection of adenoviruses G6PD (PA500+Ad-G6PD) and 500μM palmitate with infection of empty vector adenoviruses (PA500+Ad-null). Protein expression were determined by Western blot. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA500 versus cells with BSA. # denotes P < 0.05 for cells treated with PA500+Ad-G6PD versus cells with PA500+Ad-null.

    Techniques Used: Infection, Plasmid Preparation, Western Blot, Standard Deviation, Expressing

    A Overexpressing G6PD increased NADPH level in podocytes exposed to high glucose. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), high glucose with infection of adenoviruses G6PD (HG+Ad-G6PD) and high glucose with infection of empty vector adenoviruses (HG+Ad-null). NADPH was measured by a colorimetric method according to the manufacturer’s instructions. Shown are average values with standard deviation (s.d.) of triplicated experiments. ** denotes P < 0.01 for cells treated with HG versus cells with NG. # denotes P < 0.05 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. B the decreased GSH/GSSG in podocytes exposed to high glucose was ameliorated by overexpressing G6PD. Podocytes were treated as described in (A) and GSH/GSSG was measured by a spectrophotometric method following the manufacturer’s instructions. Shown are average values with standard deviation (s.d.) of triplicated experiments. ** denotes P < 0.01 for cells treated with HG versus cells with NG. ## denotes P < 0.01 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. C Elevation of G6PD protein expression reduced the accumulation of ROS in podocytes exposed to high glucose. Podocytes were treated as described in . ROS accumulation was measured with the cell-permeable, oxidation-sensitive dye CM-H 2 DCFDA. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG. ## denotes P < 0.01 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null.
    Figure Legend Snippet: A Overexpressing G6PD increased NADPH level in podocytes exposed to high glucose. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), high glucose with infection of adenoviruses G6PD (HG+Ad-G6PD) and high glucose with infection of empty vector adenoviruses (HG+Ad-null). NADPH was measured by a colorimetric method according to the manufacturer’s instructions. Shown are average values with standard deviation (s.d.) of triplicated experiments. ** denotes P < 0.01 for cells treated with HG versus cells with NG. # denotes P < 0.05 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. B the decreased GSH/GSSG in podocytes exposed to high glucose was ameliorated by overexpressing G6PD. Podocytes were treated as described in (A) and GSH/GSSG was measured by a spectrophotometric method following the manufacturer’s instructions. Shown are average values with standard deviation (s.d.) of triplicated experiments. ** denotes P < 0.01 for cells treated with HG versus cells with NG. ## denotes P < 0.01 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null. C Elevation of G6PD protein expression reduced the accumulation of ROS in podocytes exposed to high glucose. Podocytes were treated as described in . ROS accumulation was measured with the cell-permeable, oxidation-sensitive dye CM-H 2 DCFDA. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with NG. ## denotes P < 0.01 for cells treated with HG+Ad-G6PD versus cells with HG+Ad-null.

    Techniques Used: Infection, Plasmid Preparation, Standard Deviation, Expressing

    A Palmitate decreased the level of G6PD mRNA. Podocytes were treated with BSA, 500μM palmitate (PA500) for 24 hours. G6PD mRNA level was measured by real-time PCR. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA500 versus cells with BSA. B High glucose had no effect on G6PD mRNA level. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), or normal glucose supplemented with mannitol (NG+M) for 72 hours. Real-time PCR was used to analyze G6PD mRNA abundance. Shown are average values with standard deviation (s.d.) of triplicated experiments. C The degradation of G6PD induced by high glucose was in a time-dependent manner. Podocytes were incubated with high glucose (HG, 25mM glucose) for the indicated time, and cell lysates were subjected to G6PD and β-actin immunoblotting. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG for 72 hours versus cells incubated in normal glucose. D The decreased G6PD protein level in high glucose was largely rescued by MG132. Podocytes were cultured with normal glucose (NG, 5.6mM glucose), normal glucose supplemented with 0.5μM MG132 (NG+MG132), high glucose (HG, 25mM glucose) or high glucose supplemented with 0.5μM MG132 (HG+MG132). Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with HG+MG132. E G6PD was ubiquitylated. HEK293T cells were transfected with indicated plasmids with or without 20μM MG132 for 12 hours. Cell lysates were subjected to immunoprecipitation (IP) with anti-Flag M2 beads. The precipitates were probed using His and Flag antibodies. Input cell lysates were subjected to Flag and GAPDH immunoblotting.
    Figure Legend Snippet: A Palmitate decreased the level of G6PD mRNA. Podocytes were treated with BSA, 500μM palmitate (PA500) for 24 hours. G6PD mRNA level was measured by real-time PCR. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with PA500 versus cells with BSA. B High glucose had no effect on G6PD mRNA level. Podocytes were treated with normal glucose (NG, 5.6mM glucose), high glucose (HG, 25mM glucose), or normal glucose supplemented with mannitol (NG+M) for 72 hours. Real-time PCR was used to analyze G6PD mRNA abundance. Shown are average values with standard deviation (s.d.) of triplicated experiments. C The degradation of G6PD induced by high glucose was in a time-dependent manner. Podocytes were incubated with high glucose (HG, 25mM glucose) for the indicated time, and cell lysates were subjected to G6PD and β-actin immunoblotting. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG for 72 hours versus cells incubated in normal glucose. D The decreased G6PD protein level in high glucose was largely rescued by MG132. Podocytes were cultured with normal glucose (NG, 5.6mM glucose), normal glucose supplemented with 0.5μM MG132 (NG+MG132), high glucose (HG, 25mM glucose) or high glucose supplemented with 0.5μM MG132 (HG+MG132). Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with HG versus cells with HG+MG132. E G6PD was ubiquitylated. HEK293T cells were transfected with indicated plasmids with or without 20μM MG132 for 12 hours. Cell lysates were subjected to immunoprecipitation (IP) with anti-Flag M2 beads. The precipitates were probed using His and Flag antibodies. Input cell lysates were subjected to Flag and GAPDH immunoblotting.

    Techniques Used: Real-time Polymerase Chain Reaction, Standard Deviation, Incubation, Western Blot, Cell Culture, Transfection, Immunoprecipitation

    A Flag-G6PD plasmid was expressed in HEK293T cells. Flag-G6PD plasmid was transfected into HEK293T cells for 48 hours. Cells lysates were subjected to Flag and GAPDH immunoblotting. B His-Ub plasmid was verified in HEK293T cells. HEK293T cells were transfected with His-Ub plasmid for 48 hours. Protein level of His was analyzed by Western blot with anti-His antibody. C HA-VHL plasmid was expressed in HEK293T. HEK293T cells were transfected with HA-VHL plasmid for 48 hours and cells lysates were subjected to HA and GAPDH immunoblotting.
    Figure Legend Snippet: A Flag-G6PD plasmid was expressed in HEK293T cells. Flag-G6PD plasmid was transfected into HEK293T cells for 48 hours. Cells lysates were subjected to Flag and GAPDH immunoblotting. B His-Ub plasmid was verified in HEK293T cells. HEK293T cells were transfected with His-Ub plasmid for 48 hours. Protein level of His was analyzed by Western blot with anti-His antibody. C HA-VHL plasmid was expressed in HEK293T. HEK293T cells were transfected with HA-VHL plasmid for 48 hours and cells lysates were subjected to HA and GAPDH immunoblotting.

    Techniques Used: Plasmid Preparation, Transfection, Western Blot

    A Human G6PD interacted with the E3 ubiquitin ligase VHL in Y2H system. Using human G6PD as bait, VHL was identified as an interacting protein with G6PD in yeast. G6PD and VHL were co-transformed into yeast strain Mav203-activated expression of β-galactosidase. AD, Activation Domain; BD, Binding Domain; SD-2, deficient in Leucine and Tryptophan; SD-4, deficient in Leucine, Tryptophan, Histidine, and Uracil. B G6PD interacted with VHL. Co-immunoprecipitation assay shown that tagged G6PD and VHL formed a complex in HEK293T cells. HEK293T cells were transfected with HA-VHL expression plasmid in combination with or without Flag-G6PD expression plasmid. Flag-G6PD was immunoprecipitated with anti-Flag M2 beads. The precipitates were probed using HA and Flag antibodies. C VHL negatively regulated the expression of G6PD protein. HEK293T cells were co-transfected with Flag-G6PD expression plasmid and increasing amounts of HA-VHL expression plasmid. The levels of Flag and HA were determined by Western blot with indicated antibodies. D VHL reduced G6PD protein stability. HEK293T cells were transfected with Flag-G6PD expression plasmid with or without HA-VHL expression plasmid. After 36 hours, cells were treated with CHX (100μg/ml) for the indicated time. Cell lysates were subjected to Flag, GAPDH and HA immunoblotting. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells co-transfected with Flag-G6PD and HA-VHL (Flag-G6PD + HA-VHL) versus cells transfected with Flag-G6PD. E G6PD was efficiently ubiquitylated in the presence of VHL. HEK293T cells were transfected with different combinations of plasmids as indicated. G6PD was immunoprecipitated with anti-Flag M2 beads and immunoblotted with anti-His antibody to detect ubiquitylated G6PD. F Knockdown of endogenous VHL enhanced G6PD protein abundance. HEK293T cells were transfected with scrambled siRNA (scramble), siVHL#1 and siVHL#2. The levels of endogenous G6PD and VHL were analyzed by Western blot with indicated antibodies. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with siVHL#2 versus cells with scramble.
    Figure Legend Snippet: A Human G6PD interacted with the E3 ubiquitin ligase VHL in Y2H system. Using human G6PD as bait, VHL was identified as an interacting protein with G6PD in yeast. G6PD and VHL were co-transformed into yeast strain Mav203-activated expression of β-galactosidase. AD, Activation Domain; BD, Binding Domain; SD-2, deficient in Leucine and Tryptophan; SD-4, deficient in Leucine, Tryptophan, Histidine, and Uracil. B G6PD interacted with VHL. Co-immunoprecipitation assay shown that tagged G6PD and VHL formed a complex in HEK293T cells. HEK293T cells were transfected with HA-VHL expression plasmid in combination with or without Flag-G6PD expression plasmid. Flag-G6PD was immunoprecipitated with anti-Flag M2 beads. The precipitates were probed using HA and Flag antibodies. C VHL negatively regulated the expression of G6PD protein. HEK293T cells were co-transfected with Flag-G6PD expression plasmid and increasing amounts of HA-VHL expression plasmid. The levels of Flag and HA were determined by Western blot with indicated antibodies. D VHL reduced G6PD protein stability. HEK293T cells were transfected with Flag-G6PD expression plasmid with or without HA-VHL expression plasmid. After 36 hours, cells were treated with CHX (100μg/ml) for the indicated time. Cell lysates were subjected to Flag, GAPDH and HA immunoblotting. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells co-transfected with Flag-G6PD and HA-VHL (Flag-G6PD + HA-VHL) versus cells transfected with Flag-G6PD. E G6PD was efficiently ubiquitylated in the presence of VHL. HEK293T cells were transfected with different combinations of plasmids as indicated. G6PD was immunoprecipitated with anti-Flag M2 beads and immunoblotted with anti-His antibody to detect ubiquitylated G6PD. F Knockdown of endogenous VHL enhanced G6PD protein abundance. HEK293T cells were transfected with scrambled siRNA (scramble), siVHL#1 and siVHL#2. The levels of endogenous G6PD and VHL were analyzed by Western blot with indicated antibodies. Shown are average values with standard deviation (s.d.) of triplicated experiments. * denotes P < 0.05 for cells treated with siVHL#2 versus cells with scramble.

    Techniques Used: Transformation Assay, Expressing, Activation Assay, Binding Assay, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Standard Deviation

    A The map of the 5 lysine sites for VHL mediated ubiquitylation on G6PD. HEK293T cells were transfected with Flag-G6PD, His-Ub and HA-VHL plasmids, and then ubiquitylated G6PD were immunoprecipitated with anti-Flag M2 beads. MS spectra analysis identified 5 Lys residues (K 97 , K 366 , K 403 , K 407 and K 408 ) for VHL-mediated ubiquitylation in G6PD. B K 366 and K 403 were the major sites for VHL-mediated ubiquitylation on G6PD. HEK293T cells were transfected with Flag-tagged wild-type G6PD (WT) or its mutants bearing single Lys-to-Arg substitutions at the above 5 potential ubiquitylation sites. Cell lysates were subjected to immunoprecipitation (IP) with anti-Flag M2 beads. The precipitates were probed using His and Flag antibodies. C Mutants with both K 366 and K 403 sites (K 366 + 403R ) largely abolished the ubiquitylation of G6PD. HEK293T cells were transfected with Flag-tagged wild-type G6PD (WT) or G6PD mutants bearing single or two Lys-to-Arg substitutions at K 366 and K 403 sites.
    Figure Legend Snippet: A The map of the 5 lysine sites for VHL mediated ubiquitylation on G6PD. HEK293T cells were transfected with Flag-G6PD, His-Ub and HA-VHL plasmids, and then ubiquitylated G6PD were immunoprecipitated with anti-Flag M2 beads. MS spectra analysis identified 5 Lys residues (K 97 , K 366 , K 403 , K 407 and K 408 ) for VHL-mediated ubiquitylation in G6PD. B K 366 and K 403 were the major sites for VHL-mediated ubiquitylation on G6PD. HEK293T cells were transfected with Flag-tagged wild-type G6PD (WT) or its mutants bearing single Lys-to-Arg substitutions at the above 5 potential ubiquitylation sites. Cell lysates were subjected to immunoprecipitation (IP) with anti-Flag M2 beads. The precipitates were probed using His and Flag antibodies. C Mutants with both K 366 and K 403 sites (K 366 + 403R ) largely abolished the ubiquitylation of G6PD. HEK293T cells were transfected with Flag-tagged wild-type G6PD (WT) or G6PD mutants bearing single or two Lys-to-Arg substitutions at K 366 and K 403 sites.

    Techniques Used: Transfection, Immunoprecipitation

    human g6pd  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human g6pd
    Human G6pd, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human g6pd/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human g6pd - by Bioz Stars, 2023-11
    88/100 stars

    Images

    human g6pd  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human g6pd
    Human G6pd, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human g6pd/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human g6pd - by Bioz Stars, 2023-11
    88/100 stars

    Images

    human g6pd  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human g6pd
    Human G6pd, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human g6pd/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human g6pd - by Bioz Stars, 2023-11
    88/100 stars

    Images

    human g6pd  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human g6pd
    Human G6pd, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human g6pd/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human g6pd - by Bioz Stars, 2023-11
    88/100 stars

    Images

    human g6pd  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human g6pd
    Human G6pd, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human g6pd/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human g6pd - by Bioz Stars, 2023-11
    88/100 stars

    Images

    human g6pd cdna  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human g6pd cdna
    Human G6pd Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human g6pd cdna/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human g6pd cdna - by Bioz Stars, 2023-11
    88/100 stars

    Images

    human full length g6pd cdna  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 88
      Buy from Supplier

    Structured Review

    OriGene human full length g6pd cdna
    Human Full Length G6pd Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human full length g6pd cdna/product/OriGene
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human full length g6pd cdna - by Bioz Stars, 2023-11
    88/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars

    • Image Search Results