Journal: American Journal of Translational Research

Article Title: Klotho mitigates diquat-induced myocardial injury in rats by activating Nrf2/ARE-mediated suppression of oxidative stress

doi: 10.62347/SKBR3572

Figure Lengend Snippet: Klotho alleviated DQ-induced cardiomyocyte oxidative stress. (A, B) Cell viability was examined using Cell Counting Kit-8 (CCK-8) assay. (C) Klotho protein levels were examined using western blot analysis. (D) DQ-stimulated H9c2 cardiomyocytes were treated with recombinant Klotho protein, followed by viability assessment using CCK-8 assay. (E, F) Apoptosis was examined using flow cytometry. (G, H) ROS fluorescence signal, (I) MDA level, (J) GSH-ST, (K) GSH-PX, and (L) SOD activities in H9c2 cells were assessed using ELISA kits. (M, N) Nrf2, HO-1, and NQO1 protein levels were examined using western blot analysis. Data were presented as mean ± SD. * P <0.05, ** P <0.01, compared to control group. ## P <0.01, compared to DQ group.

Article Snippet: To induce oxidative injury, cells were treated with 50 μM DQ (45422, Sigma-Aldrich, USA) for 24 h. For Klotho intervention, cells were treated with 1 μg/mL recombinant Klotho protein (1819-KL, R&D Systems, USA) [ 19 , 20 ].

Techniques: Cell Counting, CCK-8 Assay, Western Blot, Recombinant, Flow Cytometry, Fluorescence, Enzyme-linked Immunosorbent Assay, Control

Journal: American Journal of Translational Research

Article Title: Klotho mitigates diquat-induced myocardial injury in rats by activating Nrf2/ARE-mediated suppression of oxidative stress

doi: 10.62347/SKBR3572

Figure Lengend Snippet: Nrf2 inhibition abrogated the protective effects of Klotho on DQ-induced oxidative stress. DQ-stimulated H9c2 cells received recombinant Klotho protein and Nrf2 inhibitor ML385. (A, B) Nrf2, HO-1, and NQO1 protein levels were examined using western blot analysis. (C) Cell viability was examined using CCK-8 assay. (D, E) Apoptosis was examined using flow cytometry. (F, G) ROS fluorescence signal, (H) MDA level, (I) GSH-ST, (J) GSH-PX, and (K) SOD activities in H9c2 cells were assessed using ELISA kits. Data were presented as mean ± SD. ** P <0.01, compared to control group. ## P <0.01, compared to DQ group. && P <0.01, compared to DQ+Klotho group.

Article Snippet: To induce oxidative injury, cells were treated with 50 μM DQ (45422, Sigma-Aldrich, USA) for 24 h. For Klotho intervention, cells were treated with 1 μg/mL recombinant Klotho protein (1819-KL, R&D Systems, USA) [ 19 , 20 ].

Techniques: Inhibition, Recombinant, Western Blot, CCK-8 Assay, Flow Cytometry, Fluorescence, Enzyme-linked Immunosorbent Assay, Control

Journal: American Journal of Translational Research

Article Title: Klotho mitigates diquat-induced myocardial injury in rats by activating Nrf2/ARE-mediated suppression of oxidative stress

doi: 10.62347/SKBR3572

Figure Lengend Snippet: Klotho alleviated DQ-induced acute myocardial injury in rats through Nrf2/ARE activation. Rats were intragastrically administered with DQ to induce acute myocardial injury and treated with recombinant Klotho protein for 5 days. (A, B) Klotho protein expression in myocardial tissues was assessed using immunohistochemistry. (C) H&E staining of myocardial tissue of rats in each group (D, E) ROS fluorescence signal, (F) MDA level, (G) GSH-ST, (H) GSH-PX, and (I) SOD activities in H9c2 cells were assessed using ELISA kits. (J, K) Nrf2, HO-1, and NQO1 protein levels in myocardial tissues was assessed using immunohistochemistry. ** P <0.01, compared to control group. ## P <0.01, compared to DQ group.

Article Snippet: To induce oxidative injury, cells were treated with 50 μM DQ (45422, Sigma-Aldrich, USA) for 24 h. For Klotho intervention, cells were treated with 1 μg/mL recombinant Klotho protein (1819-KL, R&D Systems, USA) [ 19 , 20 ].

Techniques: Activation Assay, Recombinant, Expressing, Immunohistochemistry, Staining, Fluorescence, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE 1. Klotho protects hippocampal neurons from glutamate-induced cytotoxicity. Rat primary hippocampal neurons (A–D) and immortalized mouse hippocampal neurons (HT22 cells) (E and F) were pretreated without or with Klotho for 4 h and exposed to glutamate at concentration of 3–4 mM. Cell death was assessed 24 h later by CellTiter Glo (A, D, and E) and by lactate dehydrogenase (LDH) release to the medium (C). The accumulation of intracellular ROS was assessed using 2,7-dichlorofluorescein diacetate, 8 h after the glutamate addition, and presented as -fold increase in fluorescence units normalized to untreated control (B and F). Asterisks indicate statistical significance of Klotho-treated versus Klotho-untreated cells: *, p 0.05; **, p 0.01 by Student’s t test. Results represent the average of 3–4 independent experiments. Error bars, S.D.

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques: Concentration Assay, Fluorescence, Control

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE 2. Klotho induces the expression of Trx/Prx family members in the absence or presence of glutamate. Rat primary hippocampal neurons (A and B) and mouse HT22 cells (C) were pretreated with Klotho (KL) for 4 h, followed by an additional 24 h of treatment in the absence (A) or presence (B and C) of glutamate (Glu) (3 mM). RNA was collected for qRT-PCR, and the results were normalized to endogenous controls, as described under “Experimental Proce- dures.” A, antioxidative stress array (n 5 for each group) revealed genes significantly enhanced by treatment with Klotho.B and C, the results of the array were confirmed using specific gene primer sets. The effect of Klotho was assessed in the absence or presence of glutamate. Results are representative of three independent experiments. D, rat primary hippocampal neurons were pretreated with Klotho for 4 h, followed by an additional 24 h of treatment in the absence or presence of glutamate (3 mM), as indicated, and the lysates were collected for WB analysis. Statistical significance for treated versus untreated cells was indicated as follows. *, p 0.05; **, p 0.01 by Student’s t test; &, p 0.05 by Student’s t test, for cells treated with Klotho and glutamate versus cells treated with glutamate only. Error bars, S.D. Each experiment was repeated at least three times, and representative results are shown.

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE 3. The induction of Prx-2 is essential for the neuroprotective effect of Klotho. HT22 cells were infected with the control vector and four different constructs of shRNA. The Prx-2 expression was assessed by WB (A). HT22 cells stably infected with control vector or Prx-2 shRNA3 were pretreated with Klotho (KL) for 4 h and challenged with glutamate at the indicated concentrations. The extent of cytotoxicity was assessed 24 h later using CellTiter Glo (B). Asterisks indicate statistical significance of Klotho-treated versus Klotho-untreated samples: *, p 0.05; **, p 0.01 by Student’s t test. Error bars, S.D. Results are representative of three independent experiments. C, qRT-PCR analysis of RNA obtained from whole brain samples of WT, KL-KO, and KL-OE mice. RNA was collected,andthemRNAanalysiswasperformedusingselective gene sets and normalized to endogenous controls. Each experiment was repeated at least two times, and representative results are shown.

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques: Infection, Control, Plasmid Preparation, Construct, shRNA, Expressing, Stable Transfection, Quantitative RT-PCR

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE 4. The neuroprotective effect of Klotho is mediated mainly through the activation of Akt and partially through the activation of ERK. Rat primary hippocampal neurons were treated with Klotho for the indicated times, and the activation of Akt (A) and ERK (B) was assessed by WB. Cells were treated with 10 M LY294002 (LY) (C) and U0126 5 M (D) for 1 h before the treatment with Klotho (KL). Twenty-four hours following the treatment with Klotho, the expression of Prx-2 was assessed by WB in the absence or presence of glutamate (Glu) (3 mM). Akt and ERK phosphorylated proteins are normalized to total Akt or ERK control and presented as a -fold increase normalized to untreated control. Statistical significance for Klotho-treated versus Klotho-untreated samples was indicated as follows. *, p 0.05; **, p 0.01 by Student’s t test; &, p 0.05 for cells treated with Klotho in the presence of the inhibitor versus cells treated with Klotho only. Error bars, S.D. Results represent the average of three independent experiments, and representative blots are shown.

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques: Activation Assay, Expressing, Control

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE 5. Klotho induces the phosphorylation of Akt. Rat primary hippocampal neurons were pretreated with Klotho (KL) for 4 h followed by additional treatment for 1 h (A), 24 h (B), and 48 h (C), in the absence or presence of glutamate (Glu) (3 mM). The lysates were collected for WB analysis using primary antibody recognizing the Thr308 phosphorylated site (top panels) and Ser473 phosphorylated site (bottom panels). Phosphorylated Akt proteins are normalized to total Akt (tAkt) control and presented as -fold increase normalized to untreated control. Asterisks indicate statistical significance of Klotho-treated versus Klotho-untreated samples. *, p 0.05; **, p 0.01; #, p 0.05 for cells treated with glutamate only versus untreated cells by Student’s t test. Error bars, S.D. The average of three independent experiments and representative blots are shown.

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques: Phospho-proteomics, Control

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE 6. Klotho induces the phosphorylation of Akt and FoxO3a. Rat primary hippocampal neurons were pretreated with Klotho (KL) for 4 h followed by additional treatment for 1 and 24 h, as indicated, in the absence or presence of glutamate (Glu) (3 mM). The lysates were collected for WB analysis of the effect of Klotho on phosphorylation of FoxO3a (A), Tau (B), and GSK3 (C). All proteins are normalized to the total control (A and B) or tubulin (C) and presented as a -fold increase normalized to untreated control. Asterisks indicate statistical significance of Klotho-treated versus Klotho-untreated samples: *, p 0.05; **, p 0.01 by Student’s t test. Error bars, S.D. The average of three independent experiments and representative blots are shown.

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques: Phospho-proteomics, Control

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE 7. Hippocampal neurons obtained from KL-OE embryos are more resistant to glutamate cytotoxicity than neurons from WT embryos. A, top, WB analysis of Klotho expression in 50 g of whole brain lysate from WT and KL-OE E18 embryos (n 4 for each group). Bottom panel, WB analysis of Klotho expression in 20 l of medium containing serial dilutions of recombinant mouse Klotho ( 106 nmol), starting at a concentration of 0.4 g/ml, which corresponds to 59.2 106 nmol of Klotho (lanes a–d), 25 g of cell lysate (lanes e and f), and 20 l of medium (lanes g and h) collected from hippocampal neuronsfromWTandKL-OEembryos.BandC,mouseprimaryhippocampalneuronsfromWTandKL-OEembryosweretreatedwithglutamateattheindicated concentrations. Twenty-four (B) or eight (C) hours later, the percentage cytotoxicity was assessed using CellTiter Glo, and the accumulation of ROS was measured by 2,7-dichlorofluorescein diacetate (B and C, respectively). Error bars, S.D. Each experiment was repeated at least three times, and representative results are shown. D, qRT-PCR analysis of the expression of Prx-2 and Trxrd-1 in the presence or absence of glutamate (3 mM), 24 h after the treatment. The results are presented as -fold increase of groups. First panel, neurons from KL-OE embryos versus neurons from WT embryos in the absence of oxidative conditions; second panel, neurons from WT embryos treated with glutamate (3 mM) versus untreated; third panel, neurons from KL-OE embryos treated with glutamate(3mM)versusuntreated.E,WBanalysisoftheexpressionofPrx-2andTrxrd-1inthepresenceorabsenceofglutamate(3mM),24hafterthetreatment of hippocampal neurons obtained from WT and KL-OE embryos. Asterisks indicate statistical significance: *, p 0.05; **, p 0.01 by t test. Error bars, S.D. Each experiment was repeated three times, and representative results are shown.

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques: Expressing, Recombinant, Concentration Assay, Quantitative RT-PCR

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE 8. Klotho protects hippocampal neurons from oligomeric A-induced cytotoxicity. A–C, cytotoxicity was assessed by CellTiter Glo, 24 h after exposure to oA. Rat primary hippocampal neurons pretreated with or without Klotho (KL) for 4 h (A), mouse primary hippocampal neurons obtained from WT and KL-OE embryos (B), and HT22 cells stably transduced with control vector or Prx-2 shRNA pretreated with or without Klotho for 4 h (C) were exposed to oA (0–10 M). Cell death was assessed after 24 h by CellTiter Glo. Asterisks indicate statistical significance of Klotho-exposed versus Klotho-unexposed samples: *, p 0.05; **, p 0.01 by Student’s t test. Error bars, S.D. Each experiment was repeated at least three times, and representative results are shown. D, size exclusion chromatography for oA preparation used for the toxicity assays. The far right peaks indicate the background UV absorbance of the buffer; the peaks on the left indicate soluble aggregates of A(1–42).

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques: Stable Transfection, Transduction, Control, Plasmid Preparation, shRNA, Size-exclusion Chromatography

Journal: Journal of Biological Chemistry

Article Title: The Neuroprotective Effect of Klotho is Mediated via Regulation of Members of the Redox System

doi: 10.1074/jbc.m114.567321

Figure Lengend Snippet: FIGURE9.DiagramoftheKlothomechanismofneuroprotection.Theneu- roprotective effect of Klotho on hippocampal neurons is mediated via Akt- dependent induction of Prx-2/Trxrd-1 and is associated with inhibitory phos- phorylation of FoxO3a.

Article Snippet: Reagents—For all experiments where Klotho was added exogenously to cells, we used recombinant mouse Klotho containing the shed ectodomain of Klotho (Ala35–Lys982, recombinant mouse Klotho, catalog no. 1819-KL-050 from R&D Systems, Minneapolis, MN).

Techniques:

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Moderate aerobic exercise training decreases middle-aged induced pathologic cardiac hypertrophy by improving Klotho expression, MAPK signaling pathway, and oxidative stress status in Wistar rats

doi:

Figure Lengend Snippet: Serum levels of Klotho protein in young, middle-aged model, 4 weeks trained middle aged, and 8 weeks trained middle aged rats. Moderate exercise significantly increased Klotho in 8 weeks trained middle aged rats. ** P< 0.05 compared to young rats, ## P< 0.05 compared to middle-aged rats

Article Snippet: Finally, in the eight weeks, they reached a speed of 16 m/min, the slope of 0%, and distance traveled of 830 meters during 54 min ( ) ( Enzyme-linked immunosorbent assay (ELISA) The detection of Klotho protein in serum was performed using the corresponding ELISA kits (OKEH01598; Aviva Systems Biology Co, Ltd, CA, USA) based on standard sandwich ELISA technology.

Techniques:

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Suppression of Apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs) by Klotho Protein is Associated with Reduced Endoplasmic Reticulum Oxidative Stress and Activation of the PI3K/AKT Pathway

doi: 10.12659/MSM.911202

Figure Lengend Snippet: The effects of klotho protein treatment on the viability of human umbilical vein endothelial cells (HUVECs) treated with hydrogen peroxide (H 2 O 2 ). Results are represented as the mean ±SD. Mod – the model group treated with 200 μmol/L of hydrogen peroxide (H 2 O 2 ). The number of samples was five. # P<0.05. ## P<0.01 vs. the control. * P<0.05. ** P<0.01 vs. H 2 O 2 . @ P<0.05. @@ P<0.01 vs. 100 μg/L klotho protein. LY – LY294002, which is a PI3K inhibitor.

Article Snippet: Recombinant human klotho protein was purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Suppression of Apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs) by Klotho Protein is Associated with Reduced Endoplasmic Reticulum Oxidative Stress and Activation of the PI3K/AKT Pathway

doi: 10.12659/MSM.911202

Figure Lengend Snippet: The effects of klotho protein on human umbilical vein endothelial cell (HUVEC) functional factors, including nitric oxide (NO), and inflammatory factors, tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 after hydrogen peroxide (H 2 O 2 ) treatment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of nitric oxide (NO) ( A ), interleukin (IL)-6 ( B ), and tumor necrosis factor-α (TNF-α) ( C ) in the human umbilical vein endothelial cell (HUVEC) supernatant of each group. Results are represented as the mean ±SD. Mod – the model group, treated with 200 μmol/L of hydrogen peroxide (H 2 O 2 ). The number of samples was five. # P<0.05. ## P<0.01 vs. control. * P<0.05. ** P<0.01 vs. H 2 O 2 . @ P<0.05. @@ P<0.01 vs. 100 μg/L klotho protein. LY – LY294002, which is a PI3K inhibitor.

Article Snippet: Recombinant human klotho protein was purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Functional Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Suppression of Apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs) by Klotho Protein is Associated with Reduced Endoplasmic Reticulum Oxidative Stress and Activation of the PI3K/AKT Pathway

doi: 10.12659/MSM.911202

Figure Lengend Snippet: The effects of klotho protein treatment on reactive oxygen species (ROS) of human umbilical vein endothelial cells (HUVECs) after hydrogen peroxide (H 2 O 2 ) treatment. Dihydroethidium (DHE)-derived fluorescence probe detection at 570 nm showed that human umbilical vein endothelial cells (HUVECs) in the model group, treated with hydrogen peroxide (H 2 O 2 ) produced increased levels of reactive oxygen species (ROS), which were significantly increased compared with the untreated group. However, the ROS content of the klotho protein-treated group decreased gradually. Results are represented as the mean ±SD. Mod – the model group treated with 200 μmol/L of hydrogen peroxide (H 2 O 2 ). The number of samples was five. # P<0.05. ## P<0.01 vs. control. * P<0.05. ** P<0.01 vs. H 2 O 2 . @ P<0.05. @@ P<0.01 vs. 100 μg/L klotho protein. LY – LY294002, which is a PI3K inhibitor.

Article Snippet: Recombinant human klotho protein was purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Derivative Assay, Fluorescence, Produced, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Suppression of Apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs) by Klotho Protein is Associated with Reduced Endoplasmic Reticulum Oxidative Stress and Activation of the PI3K/AKT Pathway

doi: 10.12659/MSM.911202

Figure Lengend Snippet: ( A–D ) The effects of klotho protein treatment on the activity of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), and reduced glutathione (GSH-PX) in human umbilical vein endothelial cells (HUVECs) after hydrogen peroxide (H 2 O 2 ) treatment. Results are represented as the mean ±SD. Mod – the model group treated with 200 μmol/L of hydrogen peroxide (H 2 O 2 ). The number of samples was five. # P<0.05. ## P<0.01 vs. control. * P<0.05. ** P<0.01 vs. H 2 O 2 . @ P<0.05. @@ P<0.01 vs. 100 μg/L klotho protein. LY – LY294002, which is a PI3K inhibitor.

Article Snippet: Recombinant human klotho protein was purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Activity Assay, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Suppression of Apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs) by Klotho Protein is Associated with Reduced Endoplasmic Reticulum Oxidative Stress and Activation of the PI3K/AKT Pathway

doi: 10.12659/MSM.911202

Figure Lengend Snippet: The effects of klotho protein treatment on apoptosis of human umbilical vein endothelial cells (HUVECs) after H 2 O 2 treatment measured by flow cytometry Results are represented as the mean ±SD. Mod – the model group treated with 200 μmol/L of hydrogen peroxide (H 2 O 2 ). The number of samples was five. # P<0.05. ## P<0.01 vs. control. * P<0.05. ** P<0.01 vs. H 2 O 2 . @ P<0.05. @@ P<0.01 vs. 100 μg/L klotho protein. LY – LY294002, which is a PI3K inhibitor.

Article Snippet: Recombinant human klotho protein was purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Flow Cytometry, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Suppression of Apoptosis in Human Umbilical Vein Endothelial Cells (HUVECs) by Klotho Protein is Associated with Reduced Endoplasmic Reticulum Oxidative Stress and Activation of the PI3K/AKT Pathway

doi: 10.12659/MSM.911202

Figure Lengend Snippet: ( A–C ) The effects of klotho protein treatment on the expression of the proteins, 78 kD glucose-regulated protein (GRP78), CCAAT-enhancer-binding protein homologous protein (CHOP), and caspase-12. GRP78 and CHOP and caspase-12 protein were detected by Western blot. Results are represented as the mean ±SD. Mod – the model group treated with 200 μmol/L of hydrogen peroxide (H 2 O 2 ). The number of samples was five. # P<0.05. ## P<0.01 vs. control. * P<0.05. ** P<0.01 vs. H 2 O 2 . @ P<0.05. @@ P<0.01 vs. 100 μg/L klotho protein. LY – LY294002, which is a PI3K inhibitor.

Article Snippet: Recombinant human klotho protein was purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Expressing, Binding Assay, Western Blot, Control