Journal: Oxidative Medicine and Cellular Longevity

Article Title: Buyang Huanwu Decoction Enhances Revascularization via Akt/GSK3 β /NRF2 Pathway in Diabetic Hindlimb Ischemia

doi: 10.1155/2021/1470829

Figure Lengend Snippet: BHD activates NRF2 and promotes the phosphorylation of AKT/GSK3 β during revascularization. The protein expression and quantitative analysis of NRF2 in nuclear and the phosphorylation and quantitative analysis of Akt and GSK3 β of tissue in ischemic area after 14 days of BHD treatment ( n = 4). Mean ± SD. ∗ P < 0.05, compared with HLI+NS group.

Article Snippet: BHD consists of seven herbal medicines which include Huang Qi, Dang Gui, Chi Shao, Chuan Xiong, Tao Ren, Hong Hua, and Di Long, with the ratio of 120 : 6 : 5 : 3 : 3 : 3 : 3, and the specific formulation was described previously [ ]; CD31 polyclonal antibody (ab28364) and VEGF polyclonal antibody (ab46154) were purchased from Abcam (UK); NAD(P)H dehydrogenase quinone 1 (NQO-1) polyclonal antibody (DF6437) was purchased from Affinity Biosciences (USA); heme oxygenase-1 (HO-1) polyclonal antibody (10701-1-AP), catalase polyclonal antibody (21260-1-AP), and histone H3 (17168-1-AP) were purchased from Proteintech (CHN); NRF2 monoclonal antibody (12721), Akt (4691) and phospho-Akt monoclonal antibody (4060), glycogen synthase kinase-3 beta (GSK3 β ) (12456) and phospho-GSK3 β (5558) monoclonal antibody, interleukin-6 (IL-6) monoclonal antibody (12912), interleukin-1beta (IL-1 β ) monoclonal antibody (12242), tumor necrosis factor- (TNF-) α monoclonal antibody (11948), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody (5174) were purchased from Cell Signaling Technology (USA).

Techniques: Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Buyang Huanwu Decoction Enhances Revascularization via Akt/GSK3 β /NRF2 Pathway in Diabetic Hindlimb Ischemia

doi: 10.1155/2021/1470829

Figure Lengend Snippet: BHD promotes revascularization of hindlimb of HLI mice via AKT/GSK3 β /NRF2 signal pathway. (a) The mRNA expression of NRF2 after injection of Ad-NRF2-shRNA ( n = 4). (b, c) The tissue perfusion of hindlimb in HLI mice after 0, 7, and 14 days of BHD treatment ( n = 4). (d–f) The protein expression and quantitative analysis of NRF2, VEGFA, TNF- α , IL-1 β , IL-6, CAT, NQO-1, and HO-1 and the phosphorylation and quantitative analysis of Akt and GSK3 β in ischemic area after 14 days of BHD treatment ( n = 4). Mean ± SD. NS, P > 0.05; ∗ P < 0.05, compared with HLI+BHD+sh-NRF2 group.

Article Snippet: BHD consists of seven herbal medicines which include Huang Qi, Dang Gui, Chi Shao, Chuan Xiong, Tao Ren, Hong Hua, and Di Long, with the ratio of 120 : 6 : 5 : 3 : 3 : 3 : 3, and the specific formulation was described previously [ ]; CD31 polyclonal antibody (ab28364) and VEGF polyclonal antibody (ab46154) were purchased from Abcam (UK); NAD(P)H dehydrogenase quinone 1 (NQO-1) polyclonal antibody (DF6437) was purchased from Affinity Biosciences (USA); heme oxygenase-1 (HO-1) polyclonal antibody (10701-1-AP), catalase polyclonal antibody (21260-1-AP), and histone H3 (17168-1-AP) were purchased from Proteintech (CHN); NRF2 monoclonal antibody (12721), Akt (4691) and phospho-Akt monoclonal antibody (4060), glycogen synthase kinase-3 beta (GSK3 β ) (12456) and phospho-GSK3 β (5558) monoclonal antibody, interleukin-6 (IL-6) monoclonal antibody (12912), interleukin-1beta (IL-1 β ) monoclonal antibody (12242), tumor necrosis factor- (TNF-) α monoclonal antibody (11948), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody (5174) were purchased from Cell Signaling Technology (USA).

Techniques: Expressing, Injection, shRNA

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Buyang Huanwu Decoction Enhances Revascularization via Akt/GSK3 β /NRF2 Pathway in Diabetic Hindlimb Ischemia

doi: 10.1155/2021/1470829

Figure Lengend Snippet: BHD activates NRF2 through Akt/GSK3 β signal pathway. After injection of inhibitor, the protein expression and quantitative analysis of NRF2 in nuclear and the phosphorylation and quantitative analysis of Akt and GSK3 β in ischemic area ( n = 4). Mean ± SD. ∗ P < 0.05, compared with HLI+BHD group.

Article Snippet: BHD consists of seven herbal medicines which include Huang Qi, Dang Gui, Chi Shao, Chuan Xiong, Tao Ren, Hong Hua, and Di Long, with the ratio of 120 : 6 : 5 : 3 : 3 : 3 : 3, and the specific formulation was described previously [ ]; CD31 polyclonal antibody (ab28364) and VEGF polyclonal antibody (ab46154) were purchased from Abcam (UK); NAD(P)H dehydrogenase quinone 1 (NQO-1) polyclonal antibody (DF6437) was purchased from Affinity Biosciences (USA); heme oxygenase-1 (HO-1) polyclonal antibody (10701-1-AP), catalase polyclonal antibody (21260-1-AP), and histone H3 (17168-1-AP) were purchased from Proteintech (CHN); NRF2 monoclonal antibody (12721), Akt (4691) and phospho-Akt monoclonal antibody (4060), glycogen synthase kinase-3 beta (GSK3 β ) (12456) and phospho-GSK3 β (5558) monoclonal antibody, interleukin-6 (IL-6) monoclonal antibody (12912), interleukin-1beta (IL-1 β ) monoclonal antibody (12242), tumor necrosis factor- (TNF-) α monoclonal antibody (11948), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody (5174) were purchased from Cell Signaling Technology (USA).

Techniques: Injection, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Salidroside Attenuates High-Fat Diet-Induced Nonalcoholic Fatty Liver Disease via AMPK-Dependent TXNIP/NLRP3 Pathway

doi: 10.1155/2018/8597897

Figure Lengend Snippet: Salidroside (SAL) increases phosphorylation of AMPK, ACC, Akt, and GSK3 β and suppresses activation of NLRP3 inflammasome in HFD mice. After treatment with SAL (100 mg·kg −1 ·d −1 ) for 8 weeks, liver tissues were obtained from RD and HFD mice. For the assessment of insulin sensitivity in vivo , mice were stimulated with 0.75 mU·g −1 insulin for 10 min after a 12 h fast before sacrificed. The phosphorylation of AMPK, ACC (a), Akt, and GSK3 β (b) and the activation of NLRP3 inflammasome (c) were analyzed by immunoblot. † P < 0.05, †† P < 0.01 versus RD mice treated with vehicle; ∗ P < 0.05, ∗∗ P < 0.01 versus HFD mice treated with vehicle. Values are means ± s.e.m. ( n = 3).

Article Snippet: The following primary antibodies were used: anti-IL-1 β (number 12242), anti-AMPK (number 2532), anti-phospho-AMPK Thr 172 (number 2535), anti-phospho-GSK3 β Ser 9 (number 5558), anti-acetyl coenzyme A carboxylase (ACC, number 3676), anti-ACC Ser 79 (number 3661) (Cell Signaling Technology, Beverly, MA, USA), anti-phospho-Akt Ser 473 (number 2118-1) (Epitomics, Burlingame, CA, USA), anti-caspase-1 (number 22915-1-AP), anti-NLRP3 (number 19771-1-AP), anti-GSK3 β (number 22104-1-AP) (Proteintech Group, Chicago, IL, USA), anti-TXNIP (number sc-67134) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-Akt (number A0001)(ABclonal Biotech Co. Ltd., Cambridge, MA, USA).

Techniques: Activation Assay, In Vivo, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Salidroside Attenuates High-Fat Diet-Induced Nonalcoholic Fatty Liver Disease via AMPK-Dependent TXNIP/NLRP3 Pathway

doi: 10.1155/2018/8597897

Figure Lengend Snippet: Salidroside (SAL) improves insulin sensitivity and suppresses NLRP3 inflammasome activation in hepatocytes. After cultured overnight in the serum-free medium which contains normal glucose (NG, 5.5 mM), primary mouse hepatocytes were incubated in the serum-free medium which contains 30 mM glucose and 100 nM insulin (HG) and treated with 10 μ M SAL for the indicated periods of time (0–72 h), or treated with SAL (0.1, 1, and 10 μ M) for 72 h. For assessment of insulin sensitivity in vitro , hepatocytes were treated as indicated, the medium was removed, and cells were incubated in fresh serum-free DMEM containing insulin (10 nM) for 20 min before protein samples were extracted. Protein sample were extracted from hepatocytes or supernatant (SN). The phosphorylation of Akt and GSK3 β (a, d) and the activation of NLRP3 inflammasome (c, f) were analyzed by immunoblot. The supernatant IL-1 β concentration (b, e) was measured by ELISA method. † P < 0.05, †† P < 0.01 versus NG; ∗ P < 0.05, ∗∗ P < 0.01 versus HG. Values are means ± s.e.m. ((a), (c), (d), and (f): n = 4; (b) and (e): n = 3).

Article Snippet: The following primary antibodies were used: anti-IL-1 β (number 12242), anti-AMPK (number 2532), anti-phospho-AMPK Thr 172 (number 2535), anti-phospho-GSK3 β Ser 9 (number 5558), anti-acetyl coenzyme A carboxylase (ACC, number 3676), anti-ACC Ser 79 (number 3661) (Cell Signaling Technology, Beverly, MA, USA), anti-phospho-Akt Ser 473 (number 2118-1) (Epitomics, Burlingame, CA, USA), anti-caspase-1 (number 22915-1-AP), anti-NLRP3 (number 19771-1-AP), anti-GSK3 β (number 22104-1-AP) (Proteintech Group, Chicago, IL, USA), anti-TXNIP (number sc-67134) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-Akt (number A0001)(ABclonal Biotech Co. Ltd., Cambridge, MA, USA).

Techniques: Activation Assay, Cell Culture, Incubation, In Vitro, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Salidroside Attenuates High-Fat Diet-Induced Nonalcoholic Fatty Liver Disease via AMPK-Dependent TXNIP/NLRP3 Pathway

doi: 10.1155/2018/8597897

Figure Lengend Snippet: Salidroside (SAL) protects hepatocytes against ROS overproduction. After cultured overnight in the serum-free medium which contains normal glucose (NG, 5.5 mM), hepatocytes were incubated in the serum-free medium which contains 30 mM glucose and 100 nM insulin (HG) and treated with 10 μ M SAL for the indicated periods of time (0–72 h), or treated with SAL (0.1, 1, 10 μ M) and N-acetylcysteine (NAC, 5 mM) for 72 h. The ROS levels were detected using DCF-DA and MitoSOX as indicated (a–d). The Oil Red O staining (e) and lipid content analysis (f) were performed. Scale bar = 200 μ m. Protein sample was extracted from hepatocytes or supernatant (SN). For assessment of insulin sensitivity in vitro , hepatocytes were treated as indicated, the medium was removed, and cells were incubated in fresh serum-free DMEM containing insulin (10 nM) for 20 min before protein samples were extracted. The phosphorylation of AMPK, ACC (g), Akt, and GSK3 β (h) and the activation of NLRP3 inflammasome (j) were analyzed by immunoblot. The supernatant IL-1 β concentration was measured by ELISA method (i). † P < 0.05, †† P < 0.01 versus NG; ∗ P < 0.05, ∗∗ P < 0.01 versus HG. Values are means ± s.e.m. ((a)–(d), (f)–(h), and (j): n = 4; (e) and (i): n = 3).

Article Snippet: The following primary antibodies were used: anti-IL-1 β (number 12242), anti-AMPK (number 2532), anti-phospho-AMPK Thr 172 (number 2535), anti-phospho-GSK3 β Ser 9 (number 5558), anti-acetyl coenzyme A carboxylase (ACC, number 3676), anti-ACC Ser 79 (number 3661) (Cell Signaling Technology, Beverly, MA, USA), anti-phospho-Akt Ser 473 (number 2118-1) (Epitomics, Burlingame, CA, USA), anti-caspase-1 (number 22915-1-AP), anti-NLRP3 (number 19771-1-AP), anti-GSK3 β (number 22104-1-AP) (Proteintech Group, Chicago, IL, USA), anti-TXNIP (number sc-67134) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-Akt (number A0001)(ABclonal Biotech Co. Ltd., Cambridge, MA, USA).

Techniques: Cell Culture, Incubation, Staining, In Vitro, Activation Assay, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: IRS-2/Akt/GSK-3 β /Nrf2 Pathway Contributes to the Protective Effects of Chikusetsu Saponin IVa against Lipotoxicity

doi: 10.1155/2021/8832318

Figure Lengend Snippet: CHS induced the activation of IRS-2/Akt/GSK-3 β signaling in PA-treated β TC3 cells. After being treated with CHS (24 h) and/or PA (another 24 h), the phosphorylation levels of IRS-2 (a), Akt (b), and GSK-3 β (c) were examined. The columns and errors bars are presented as means ± SEM ( n ≥ 5). ## P < 0.01 compared with the CON group. ∗∗ P < 0.01 compared with the PA group. △△ P < 0.01 and △ P < 0.05 compared with the CHS (20 μ M) group.

Article Snippet: Rabbit anti-Nrf2 (#12721, 1 : 1000, Cell Signaling Technology), rabbit anti-IRS-2 (#3089, 1 : 1000, Cell Signaling Technology), rabbit anti-Akt (#9272, 1 : 1000, Cell Signaling Technology), rabbit anti-P-Akt (#4060, 1 : 2000, Cell Signaling Technology), rabbit anti-GSK-3 β (#12456, 1 : 1000, Cell Signaling Technology), rabbit anti-P-GSK-3 β (Ser9) (#5558, 1 : 1000, Cell Signaling Technology), rabbit anti-PDX-1 (#5679, 1 : 1000, Cell Signaling Technology), rabbit anti-SOD1 (#2770, 1 : 1000, Cell Signaling Technology), rabbit anti- β -actin (#4970, 1 : 1000, Cell Signaling Technology), and rabbit anti-Lamin B (#12255, 1 : 1000, Cell Signaling Technology).

Techniques: Activation Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: IRS-2/Akt/GSK-3 β /Nrf2 Pathway Contributes to the Protective Effects of Chikusetsu Saponin IVa against Lipotoxicity

doi: 10.1155/2021/8832318

Figure Lengend Snippet: CHS protected against oxidative stress of pancreatic β TC3 cells from lipotoxicity through GSK-3 β and the relationships between Nrf2 and GSK-3 β . β TC3 cells were treated with CHS in the presence or absence of LiCl (GSK-3 β inhibitor) for 24 h and then subjected to PA for another 24 h. Effects of LiCl (GSK-3 β inhibitor) on CHS-induced GSK-3 β phosphorylation and PDX-1 expression levels were measured (a). Immunoblot analysis showed the protein expression of Nrf2 in β TC3 cells pretreated with CHS (24 h) with or without PA (24 h) (b). Effects of LiCl on Nrf2 transcriptional activity (c), ROS (d), SOD (e), cell viability (f), and insulin levels (g) were measured after different treatments. The columns and errors bars are presented as means ± SEM ( n ≥ 5). ## P < 0.01 compared with the CON group, ∗∗ P < 0.01 compared with the PA group, and && P < 0.01 compared with the CHS treatment group. (h) Potential mechanism underlying the protective effects of CHS on lipotoxicity-induced cell injuries.

Article Snippet: Rabbit anti-Nrf2 (#12721, 1 : 1000, Cell Signaling Technology), rabbit anti-IRS-2 (#3089, 1 : 1000, Cell Signaling Technology), rabbit anti-Akt (#9272, 1 : 1000, Cell Signaling Technology), rabbit anti-P-Akt (#4060, 1 : 2000, Cell Signaling Technology), rabbit anti-GSK-3 β (#12456, 1 : 1000, Cell Signaling Technology), rabbit anti-P-GSK-3 β (Ser9) (#5558, 1 : 1000, Cell Signaling Technology), rabbit anti-PDX-1 (#5679, 1 : 1000, Cell Signaling Technology), rabbit anti-SOD1 (#2770, 1 : 1000, Cell Signaling Technology), rabbit anti- β -actin (#4970, 1 : 1000, Cell Signaling Technology), and rabbit anti-Lamin B (#12255, 1 : 1000, Cell Signaling Technology).

Techniques: Expressing, Western Blot, Activity Assay

Journal: BioMed Research International

Article Title: Human Schlafen 5 Inhibits Proliferation and Promotes Apoptosis in Lung Adenocarcinoma via the PTEN/PI3K/AKT/mTOR Pathway

doi: 10.1155/2021/6628682

Figure Lengend Snippet: SLFN5 regulates p-AKT/GSK-3 β / β -catenin and p-AKT/mTOR pathways via transcriptional regulation of PTEN mRNA. (a) Western blot analysis of protein expression of p-AKT, GSK-3 β , β -catenin, mTOR, and PTEN in A549 cells stably transfected with ShCtrl, ShSLFN5, Vector, or SLFN5. (b) Real-time PCR analysis of PTEN in A549 cells stably transfected with ShCtrl, ShSLFN5, Vector, or SLFN5. (c) Immunohistochemistry and H&E staining of SLFN5 and PTEN in lung cancer samples. Red arrowheads indicate cancer cells, and black arrowheads indicate lymphocytes in the cancer stroma. Scale bars, 100 μ M. Positive expression analyses of PTEN and SLFN5 were shown as mean ± S.D.

Article Snippet: PTEN (D4.3) mAb (Cat # 9188S), Phospho-AKT (S473) (D9E) mAb (Cat # 4060S), TCF1/TCF7 (C63D9) mAb (Cat # 2203S), Phospho-mTOR (Ser2448) (D9C2) mAb (Cat # 5536T), mTOR (7C10) mAb (Cat # 2983T), Phospho-GSK-3-beta (S9) (D85E12) mAb (Cat # 5558S), β -catenin (D10A8) mAb (Cat #8480), caspase-3 (8G10) mAb (Cat #9665), cleaved caspase-3 (Asp175) (5A1E) mAb (Cat #9664), Bax (D2E11) mAb (Cat #5023), cyclin D1 antibody (Cat #2922), and actin antibody are all from Cell Signaling Technology.

Techniques: Western Blot, Expressing, Stable Transfection, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Immunohistochemistry, Staining

Journal: Stem Cells International

Article Title: Iron Overload Impairs Bone Marrow Mesenchymal Stromal Cells from Higher-Risk MDS Patients by Regulating the ROS-Related Wnt/ β -Catenin Pathway

doi: 10.1155/2020/8855038

Figure Lengend Snippet: The mechanisms involved in IO damaging MSCs of MDS patients. (a) Compared with the NIO MDS group and the control group, the apoptosis of MSCs significantly increased in the IO MDS group ( p < 0.01). (b) Compared with the other groups, the level of ROS significantly increased in MSCs of the IO MDS group ( p < 0.01). (c–d) The gene expressions of caspase3 and β -catenin were significantly upregulated in MSCs of the IO MDS group compared with those of the other groups. (e) The protein of p-AKT in IO MDS MSCs was downregulated, while caspase3 was upregulated. The protein of β -catenin in IO MDS MSCs was upregulated, while GSK-3 β was downregulated. Compared with the control group: ∗ indicates p < 0.05; ∗∗ indicates p < 0.01; ∗∗∗ indicates p < 0.001. Compared with the NIO MDS group: # indicates p < 0.05; ## indicates p < 0.01.

Article Snippet: NC membranes were blocked with 5% BSA for 1 h at room temperature (RT), and then incubated overnight at 4°C with the following primary antibodies: anti-AKT (#4691S), anti-p-AKT (#9271S), anti-GSK-3 β (#12456S), anti-p-GSK-3 β (#5558S), anti- β -catenin (#8480T), anti-caspase3 (#9668S), and anti-GAPDH (#5174T) (Cell Signaling Technology, USA).

Techniques:

Journal: Stem Cells International

Article Title: Iron Overload Impairs Bone Marrow Mesenchymal Stromal Cells from Higher-Risk MDS Patients by Regulating the ROS-Related Wnt/ β -Catenin Pathway

doi: 10.1155/2020/8855038

Figure Lengend Snippet: The effects of antioxidant or iron chelation treatments on IO MDS MSCs. (a) The levels of ROS in IO MDS MSCs were significantly decreased after the treatments with NAC or DFO. (b) The apoptosis of IO MDS MSCs was significantly decreased after the treatments with NAC or DFO. (c) The gene expressions of caspase3 and β -catenin in IO MDS MSCs were significantly downregulated after the treatments with NAC or DFO. (d) The protein of p-AKT in IO MDS MSCs was upregulated after the treatments with NAC or DFO, while caspase3 was downregulated. The protein of β -catenin in IO MDS MSCs was downregulated after the treatments with NAC or DFO, while GSK-3 β was upregulated. Compared with the IO MDS group: & indicates p < 0.05; && indicates p < 0.01.

Article Snippet: NC membranes were blocked with 5% BSA for 1 h at room temperature (RT), and then incubated overnight at 4°C with the following primary antibodies: anti-AKT (#4691S), anti-p-AKT (#9271S), anti-GSK-3 β (#12456S), anti-p-GSK-3 β (#5558S), anti- β -catenin (#8480T), anti-caspase3 (#9668S), and anti-GAPDH (#5174T) (Cell Signaling Technology, USA).

Techniques:

Journal: Stem Cells International

Article Title: Iron Overload Impairs Bone Marrow Mesenchymal Stromal Cells from Higher-Risk MDS Patients by Regulating the ROS-Related Wnt/ β -Catenin Pathway

doi: 10.1155/2020/8855038

Figure Lengend Snippet: The characteristics and functions of MSCs between IO MDS and IO MDS/AML patients. (a) There was no difference in morphology of MSCs between IO MDS and MDS/AML patients. (b) The level of ROS in the IO MDS/AML group was significantly higher than that in the IO MDS group ( p < 0.05). (c) The apoptosis of MSCs in the IO MDS/AML group was significantly higher than that in the IO MDS group. (d) The gene expressions of caspase3 and β -catenin in MSCs in the IO MDS/AML group were significantly upregulated compared with those in the IO MDS group, with a significantly downregulated gene expression of CXCL12. (e) The protein levels of p-AKT and GSK-3 β in the IO MDS/AML group were downregulated, with upregulated protein levels of caspase3 and β -catenin. Compared with the IO MDS group: ∗ indicates p < 0.05.

Article Snippet: NC membranes were blocked with 5% BSA for 1 h at room temperature (RT), and then incubated overnight at 4°C with the following primary antibodies: anti-AKT (#4691S), anti-p-AKT (#9271S), anti-GSK-3 β (#12456S), anti-p-GSK-3 β (#5558S), anti- β -catenin (#8480T), anti-caspase3 (#9668S), and anti-GAPDH (#5174T) (Cell Signaling Technology, USA).

Techniques: Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TRPV1 Modulator Ameliorates Alzheimer-Like Amyloid- β Neuropathology via Akt/Gsk3 β -Mediated Nrf2 Activation in the Neuro-2a/APP Cell Model

doi: 10.1155/2022/1544244

Figure Lengend Snippet: PcActx-induced Nrf2 activation and A β downregulation were mediated by the Akt/Gsk3 β pathway. (a–c) PcActx peptide promoted the phosphorylation of Akt (Ser 473) and Gsk3 β (Ser 9). (d–h) An Akt inhibitor suppressed the PcActx-induced upregulation of Nrf2, NQO1, and HO-1, as well as the downregulation of Keap1. (i, j ) The reductions of APP, A β 1-40, and A β 1-42 were counteracted by pharmacological inhibition of Akt/Gsk3 β signaling. Data are expressed as the mean ± SD of three or four independent experiments, in triplicate. # p < 0.05 vs. N2a/APP group and ## p < 0.01 vs. N2a/APP group. ∗ p < 0.05 vs. PcActx-treated group and ∗∗ p < 0.01 vs. PcActx-treated group.

Article Snippet: Antibodies against A β 1-40 (Cat. No.: 12990), A β 1-42 (Cat. No.: 14974), PSEN1 (Cat. No.: 5643), PSEN2 (Cat. No.: 9979), p-CaMKII (Thr 286, Cat. No.:12716), CaMKII (Cat. No.: 11945), CaMKIV (Cat. No.: 4032), Nrf2 (Cat. No.: 12721), Keap1 (Cat. No.: 8047), NQO1 (Cat. No.: 62262), HO-1 (Cat. No.: 43966), p-Akt (Ser 473, Cat. No.: 4060), Akt (Cat. No.: 4691), p-Gsk3 β (Ser 9, Cat. No.: 5558), Gsk3 β (Cat. No.: 9315), GAPDH (Cat. No.: 2118), and lamin B (Cat. No.: 13435) and Alexa-Fluor-488 conjugated anti-rabbit IgG secondary antibody (Cat. No.: 4412) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Activation Assay, Inhibition