rabbit anti glycogen synthase kinase 3 beta  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti glycogen synthase kinase 3 beta
    Experimental scheme for this study. After gene identification at 1 month of age, 3-month-old male WT and 3×Tg-AD mice were randomly assigned to four groups with 10 animals each and then intragastrically administered either ICA or vehicle for 5 months (WT + vehicle, WT + ICA, 3×Tg-AD + vehicle, 3×Tg-AD + ICA groups). After performing behavior tests, the mice were euthanized. The cerebral cortexes were evaluated using HE and Nissl staining, immunofluorescent staining, and western blot assays to determine the above disease indicators. 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; Aβ: beta-amyloid protein; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; <t>GSK3β:</t> glycogen synthase <t>kinase</t> <t>3</t> beta; HE: hematoxylin and eosin; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; NeuN: neuronal nuclear antigen; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PSD95: postsynaptic density protein 95; WT: wild-type.
    Rabbit Anti Glycogen Synthase Kinase 3 Beta, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Icariin ameliorates memory deficits through regulating brain insulin signaling and glucose transporters in 3×Tg-AD mice"

    Article Title: Icariin ameliorates memory deficits through regulating brain insulin signaling and glucose transporters in 3×Tg-AD mice

    Journal: Neural Regeneration Research

    doi: 10.4103/1673-5374.344840

    Experimental scheme for this study. After gene identification at 1 month of age, 3-month-old male WT and 3×Tg-AD mice were randomly assigned to four groups with 10 animals each and then intragastrically administered either ICA or vehicle for 5 months (WT + vehicle, WT + ICA, 3×Tg-AD + vehicle, 3×Tg-AD + ICA groups). After performing behavior tests, the mice were euthanized. The cerebral cortexes were evaluated using HE and Nissl staining, immunofluorescent staining, and western blot assays to determine the above disease indicators. 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; Aβ: beta-amyloid protein; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; GSK3β: glycogen synthase kinase 3 beta; HE: hematoxylin and eosin; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; NeuN: neuronal nuclear antigen; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PSD95: postsynaptic density protein 95; WT: wild-type.
    Figure Legend Snippet: Experimental scheme for this study. After gene identification at 1 month of age, 3-month-old male WT and 3×Tg-AD mice were randomly assigned to four groups with 10 animals each and then intragastrically administered either ICA or vehicle for 5 months (WT + vehicle, WT + ICA, 3×Tg-AD + vehicle, 3×Tg-AD + ICA groups). After performing behavior tests, the mice were euthanized. The cerebral cortexes were evaluated using HE and Nissl staining, immunofluorescent staining, and western blot assays to determine the above disease indicators. 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; Aβ: beta-amyloid protein; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; GSK3β: glycogen synthase kinase 3 beta; HE: hematoxylin and eosin; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; NeuN: neuronal nuclear antigen; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PSD95: postsynaptic density protein 95; WT: wild-type.

    Techniques Used: Staining, Western Blot, Transgenic Assay

    Effects of ICA on impaired insulin signaling in the cerebral cortex of 3×Tg-AD mice. (A) Insulin signaling: IR tyrosine autophosphorylation is stimulated by insulin and triggers IRS1 phosphorylation at tyrosine residues, which represents a positive regulatory mechanism that activates the PI3K/AKT pathway and results in the inhibition of GSK3β. However, serine phosphorylation of IRS1 at specific sites is a negative regulatory mechanism. (B) Representative expression patterns of molecules related to the insulin signaling pathway. (C) Quantification of proteins related to the insulin signaling pathway shown in (B). Protein levels were normalized to those in the WT + vehicle group. The data are presented as the means ± SEM ( n = 4–6). * P < 0.05, vs . WT + vehicle group; # P < 0.05, vs . 3×Tg-AD + vehicle group (one-way analysis of variance followed by the least significant difference test). 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; AKT: protein kinase B; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GSK3β: glycogen synthase kinase 3 beta; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PIP2: phosphatidylinositol (4,5) bisphosphate; PIP3: phosphatidylinositol (3,4,5) trisphosphate; PTEN: phosphatase and tensin homolog; WT: wild-type.
    Figure Legend Snippet: Effects of ICA on impaired insulin signaling in the cerebral cortex of 3×Tg-AD mice. (A) Insulin signaling: IR tyrosine autophosphorylation is stimulated by insulin and triggers IRS1 phosphorylation at tyrosine residues, which represents a positive regulatory mechanism that activates the PI3K/AKT pathway and results in the inhibition of GSK3β. However, serine phosphorylation of IRS1 at specific sites is a negative regulatory mechanism. (B) Representative expression patterns of molecules related to the insulin signaling pathway. (C) Quantification of proteins related to the insulin signaling pathway shown in (B). Protein levels were normalized to those in the WT + vehicle group. The data are presented as the means ± SEM ( n = 4–6). * P < 0.05, vs . WT + vehicle group; # P < 0.05, vs . 3×Tg-AD + vehicle group (one-way analysis of variance followed by the least significant difference test). 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; AKT: protein kinase B; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GSK3β: glycogen synthase kinase 3 beta; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PIP2: phosphatidylinositol (4,5) bisphosphate; PIP3: phosphatidylinositol (3,4,5) trisphosphate; PTEN: phosphatase and tensin homolog; WT: wild-type.

    Techniques Used: Inhibition, Expressing, Transgenic Assay

    Schematic diagram of the mechanism by which ICA regulates GLUTs and brain insulin signaling to ameliorate memory impairment in AD. Aβ: Amyloid-beta protein; AD: Alzheimer’s disease; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; GSK3β: glycogen synthase kinase 3 beta; G-tau: the attachment of O-linked N-acetylglucosamine (O-GlcNAc) on tau; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PIP2: phosphatidylinositol (4,5) bisphosphate; PIP3: phosphatidylinositol (3,4,5) trisphosphate; PTEN: phosphatase and tensin homolog.
    Figure Legend Snippet: Schematic diagram of the mechanism by which ICA regulates GLUTs and brain insulin signaling to ameliorate memory impairment in AD. Aβ: Amyloid-beta protein; AD: Alzheimer’s disease; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; GSK3β: glycogen synthase kinase 3 beta; G-tau: the attachment of O-linked N-acetylglucosamine (O-GlcNAc) on tau; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PIP2: phosphatidylinositol (4,5) bisphosphate; PIP3: phosphatidylinositol (3,4,5) trisphosphate; PTEN: phosphatase and tensin homolog.

    Techniques Used:

    glycogen synthase kinase 3 beta  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc glycogen synthase kinase 3 beta
    Molecular docking of core components and Wnt/β-catenin-related targets. Quercetin, luteolin, fisetin, wogonin, oroxylin A, boldine, tetrahydroalstonine, and galangin with (A) β-catenin, and (B) <t>GSK3β.</t> (C) Cluster heatmap of docking results. Numerical value indicated the binding energy ((kcal/mol).
    Glycogen Synthase Kinase 3 Beta, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Stimulation of hair growth by Tianma Gouteng decoction: Identifying mechanisms based on chemical analysis, systems biology approach, and experimental evaluation"

    Article Title: Stimulation of hair growth by Tianma Gouteng decoction: Identifying mechanisms based on chemical analysis, systems biology approach, and experimental evaluation

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2022.1073392

    Molecular docking of core components and Wnt/β-catenin-related targets. Quercetin, luteolin, fisetin, wogonin, oroxylin A, boldine, tetrahydroalstonine, and galangin with (A) β-catenin, and (B) GSK3β. (C) Cluster heatmap of docking results. Numerical value indicated the binding energy ((kcal/mol).
    Figure Legend Snippet: Molecular docking of core components and Wnt/β-catenin-related targets. Quercetin, luteolin, fisetin, wogonin, oroxylin A, boldine, tetrahydroalstonine, and galangin with (A) β-catenin, and (B) GSK3β. (C) Cluster heatmap of docking results. Numerical value indicated the binding energy ((kcal/mol).

    Techniques Used: Binding Assay

    TGD promoted hair shaft elongation in rat vibrissa follicles. (A) Photographs of rat vibrissa follicles cultured with TGD for 0, 3, 6, 9, 12 days; The double arrow area represents the measured hair shaft length. (B) Changes in length of hair shafts in the vibrissa follicles treated with TGD for 12 days. Data are presented as mean ± SEM ( n = 6). ** p < 0.01 compared with the control group. (C–E) The representative images of immunofluorescence staining in rat vibrissa follicles for β-catenin, GSK3β, LEF1 (×40), picture of magnification ×200 in white box.
    Figure Legend Snippet: TGD promoted hair shaft elongation in rat vibrissa follicles. (A) Photographs of rat vibrissa follicles cultured with TGD for 0, 3, 6, 9, 12 days; The double arrow area represents the measured hair shaft length. (B) Changes in length of hair shafts in the vibrissa follicles treated with TGD for 12 days. Data are presented as mean ± SEM ( n = 6). ** p < 0.01 compared with the control group. (C–E) The representative images of immunofluorescence staining in rat vibrissa follicles for β-catenin, GSK3β, LEF1 (×40), picture of magnification ×200 in white box.

    Techniques Used: Cell Culture, Immunofluorescence, Staining

    (A–E) The representative images and statistical graph of β-catenin, GSK3β, p-GSK3β and LEF1 in DPCs (×200), n = 3 per group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group. (F–H) Western blot analysis of Wnt/β-catenin signaling pathways related protein expression in DPCs, n = 4 per group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group.
    Figure Legend Snippet: (A–E) The representative images and statistical graph of β-catenin, GSK3β, p-GSK3β and LEF1 in DPCs (×200), n = 3 per group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group. (F–H) Western blot analysis of Wnt/β-catenin signaling pathways related protein expression in DPCs, n = 4 per group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group.

    Techniques Used: Western Blot, Expressing

    rabbit anti glycogen synthase kinase gsk 3 β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti glycogen synthase kinase gsk 3 β
    Rabbit Anti Glycogen Synthase Kinase Gsk 3 β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho glycogen synthase kinase 3 beta ser9 rabbit igg  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho glycogen synthase kinase 3 beta ser9 rabbit igg
    (A) Immunoblot and densitometric analyses of (B) total and (C) phosphorylated glycogen <t>synthase</t> <t>kinase</t> <t>3</t> beta (GSK3β), p38 MAP Kinase (p38) and p44/42 MAPK (Erk) were performed in whole cell extracts from human fibroblasts deriving from control (CONT, white bars, N = 3), mutated parkin (PARK, dark grey bars, N = 3), mutated LRRK2 (LRRK2, light grey bars, N = 3) and idiopathic PD (PD, black bars, N = 3). For the quantitation, values of total protein were normalized on the level of GAPDH of the relative sample, whereas the levels of phosphorylated form were normalized on the values of total protein. All values are expressed as mean ± SEM. * p <0.05 and ** p <0.02 vs control, # p <0.05 vs PD according to ANOVA, Tukey HSD post hoc test.
    Phospho Glycogen Synthase Kinase 3 Beta Ser9 Rabbit Igg, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Microtubule Destabilization Is Shared by Genetic and Idiopathic Parkinson’s Disease Patient Fibroblasts"

    Article Title: Microtubule Destabilization Is Shared by Genetic and Idiopathic Parkinson’s Disease Patient Fibroblasts

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0037467

    (A) Immunoblot and densitometric analyses of (B) total and (C) phosphorylated glycogen synthase kinase 3 beta (GSK3β), p38 MAP Kinase (p38) and p44/42 MAPK (Erk) were performed in whole cell extracts from human fibroblasts deriving from control (CONT, white bars, N = 3), mutated parkin (PARK, dark grey bars, N = 3), mutated LRRK2 (LRRK2, light grey bars, N = 3) and idiopathic PD (PD, black bars, N = 3). For the quantitation, values of total protein were normalized on the level of GAPDH of the relative sample, whereas the levels of phosphorylated form were normalized on the values of total protein. All values are expressed as mean ± SEM. * p <0.05 and ** p <0.02 vs control, # p <0.05 vs PD according to ANOVA, Tukey HSD post hoc test.
    Figure Legend Snippet: (A) Immunoblot and densitometric analyses of (B) total and (C) phosphorylated glycogen synthase kinase 3 beta (GSK3β), p38 MAP Kinase (p38) and p44/42 MAPK (Erk) were performed in whole cell extracts from human fibroblasts deriving from control (CONT, white bars, N = 3), mutated parkin (PARK, dark grey bars, N = 3), mutated LRRK2 (LRRK2, light grey bars, N = 3) and idiopathic PD (PD, black bars, N = 3). For the quantitation, values of total protein were normalized on the level of GAPDH of the relative sample, whereas the levels of phosphorylated form were normalized on the values of total protein. All values are expressed as mean ± SEM. * p <0.05 and ** p <0.02 vs control, # p <0.05 vs PD according to ANOVA, Tukey HSD post hoc test.

    Techniques Used: Western Blot, Quantitation Assay

    glycogen synthase kinase 3 beta  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc glycogen synthase kinase 3 beta
    PGB protected against RIR injury via the <t>Akt/GSK3β/β-catenin</t> pathway. ( A ) The results of WB showed that the expression of p-Akt, p-GSK3β, and β-catenin was downregulated after RIR, whereas PGB (30 mg/kg) pretreatment reversed these findings. ( B , D ) Representative IF images of retinal sections (40×). Scale bars : 50 µm. The ratios of RGCs in the GCL expressing p-Akt, p-GSK3β, and β-catenin were downregulated after RIR, but PGB pretreatment upregulated the proportion of positive cells. ( C ) PGB pretreatment inhibited the activity of GSK3β ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + RIR group.
    Glycogen Synthase Kinase 3 Beta, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Pregabalin Mediates Retinal Ganglion Cell Survival From Retinal Ischemia/Reperfusion Injury Via the Akt/GSK3β/β-Catenin Signaling Pathway"

    Article Title: Pregabalin Mediates Retinal Ganglion Cell Survival From Retinal Ischemia/Reperfusion Injury Via the Akt/GSK3β/β-Catenin Signaling Pathway

    Journal: Investigative Ophthalmology & Visual Science

    doi: 10.1167/iovs.63.12.7

    PGB protected against RIR injury via the Akt/GSK3β/β-catenin pathway. ( A ) The results of WB showed that the expression of p-Akt, p-GSK3β, and β-catenin was downregulated after RIR, whereas PGB (30 mg/kg) pretreatment reversed these findings. ( B , D ) Representative IF images of retinal sections (40×). Scale bars : 50 µm. The ratios of RGCs in the GCL expressing p-Akt, p-GSK3β, and β-catenin were downregulated after RIR, but PGB pretreatment upregulated the proportion of positive cells. ( C ) PGB pretreatment inhibited the activity of GSK3β ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + RIR group.
    Figure Legend Snippet: PGB protected against RIR injury via the Akt/GSK3β/β-catenin pathway. ( A ) The results of WB showed that the expression of p-Akt, p-GSK3β, and β-catenin was downregulated after RIR, whereas PGB (30 mg/kg) pretreatment reversed these findings. ( B , D ) Representative IF images of retinal sections (40×). Scale bars : 50 µm. The ratios of RGCs in the GCL expressing p-Akt, p-GSK3β, and β-catenin were downregulated after RIR, but PGB pretreatment upregulated the proportion of positive cells. ( C ) PGB pretreatment inhibited the activity of GSK3β ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + RIR group.

    Techniques Used: Expressing, Activity Assay

    The neuroprotective effect of PGB required the phosphorylation of Akt and inactivation of GSK3β. ( A , B ) The results of WB showed that MK2206 (10 nM) inhibited the phosphorylation of Akt and GSK3β stimulated by PGB. ( C , D ) Representative IF images of retinal sections (40×). Scale bars : 50 µm. MK2206 inhibited the increase in the ratio of RGCs, with p-Akt + and p-GSK3β + stimulated by PGB (30 mg/kg). ( E ) Consistent with the effect of PGB pretreatment, TWS119 (10 nM) inhibited the activity of GSK3β. ( F ) WB showed that TWS119 enhanced the expression of β-catenin ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + RIR group; *** P < 0.05 versus 30-mg/kg PGB + dimethyl sulfoxide (DMSO) + RIR group.
    Figure Legend Snippet: The neuroprotective effect of PGB required the phosphorylation of Akt and inactivation of GSK3β. ( A , B ) The results of WB showed that MK2206 (10 nM) inhibited the phosphorylation of Akt and GSK3β stimulated by PGB. ( C , D ) Representative IF images of retinal sections (40×). Scale bars : 50 µm. MK2206 inhibited the increase in the ratio of RGCs, with p-Akt + and p-GSK3β + stimulated by PGB (30 mg/kg). ( E ) Consistent with the effect of PGB pretreatment, TWS119 (10 nM) inhibited the activity of GSK3β. ( F ) WB showed that TWS119 enhanced the expression of β-catenin ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + RIR group; *** P < 0.05 versus 30-mg/kg PGB + dimethyl sulfoxide (DMSO) + RIR group.

    Techniques Used: Activity Assay, Expressing

    PGB pretreatment prevented mitochondrial impairment and retinal dysfunction by the phosphorylation of Akt and inactivation of GSK3β. ( A ) PGB (30 mg/kg) and TWS119 (10 nM) pretreatment improved the structure of mitochondria in the retina of RIR mice models; mitochondria are indicated by the yellow arrow (25,000×). Scale bars : 500 nm. ( B , D ) MK2206 (10 nM) reversed the effects of PGB and decreased the thickness of the RNFL and GCC, and TWS119 maintained the effects of PGB and reserved the increased thickness of RNFL and GCC. Scale bars : 200 µm. ( C , E ) Representative ERG waveforms in living mice. MK2206 disrupted the visual function balance of RGCs in the presence of PGB, with a decrease in the amplitudes of OPs and PhNRs. However, TWS119 maintained the normal level of the amplitudes of OPs and PhNRs ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + RIR group; *** P < 0.05 versus 30-mg/kg PGB + DMSO + RIR group.
    Figure Legend Snippet: PGB pretreatment prevented mitochondrial impairment and retinal dysfunction by the phosphorylation of Akt and inactivation of GSK3β. ( A ) PGB (30 mg/kg) and TWS119 (10 nM) pretreatment improved the structure of mitochondria in the retina of RIR mice models; mitochondria are indicated by the yellow arrow (25,000×). Scale bars : 500 nm. ( B , D ) MK2206 (10 nM) reversed the effects of PGB and decreased the thickness of the RNFL and GCC, and TWS119 maintained the effects of PGB and reserved the increased thickness of RNFL and GCC. Scale bars : 200 µm. ( C , E ) Representative ERG waveforms in living mice. MK2206 disrupted the visual function balance of RGCs in the presence of PGB, with a decrease in the amplitudes of OPs and PhNRs. However, TWS119 maintained the normal level of the amplitudes of OPs and PhNRs ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + RIR group; *** P < 0.05 versus 30-mg/kg PGB + DMSO + RIR group.

    Techniques Used:

    PGB protected primary RGCs from OGD/R injury via the Akt/GSK3β/β-catenin pathway. ( A ) IF identification of primary mice RGCs (40×). Scale bars : 50 µm. ( B ) Primary RGCs pretreated with PGB (50 µM, 100 µM, or 200 µM) for 24 hours were subjected to OGD/R. OGD/R induced a considerable decrease in cell viability, whereas PGB improved cell viability. The most significant improvement was observed at 100 µM. ( C , D ) Primary RGCs pretreated with MK2206 (5 µM) were treated with PGB (100 µM) for 24 hours prior to the onset of OGD/R. The ROS level was measured through the fluorescent probe, DCFH-DA, and the ratio of ROS + cells was measured. PGB pretreatment inhibited ROS generation, but MK2206 counteracted the effect. (40×). Scale bars : 50 µm. ( E ) Representative IF images of primary RGCs (40×). Scale bars : 50 µm. The ratios of RGCs expressing p-Akt, p-GSK3β, and β-catenin were downregulated after OGD/R, which was countered by PGB pretreatment. However, MK2206 decreased the ratios of positive cells ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + OGD/R group; *** P < 0.05 versus PGB 100 µM + OGD/R group; **** P < 0.05 versus PGB + DMSO + OGD/R group.
    Figure Legend Snippet: PGB protected primary RGCs from OGD/R injury via the Akt/GSK3β/β-catenin pathway. ( A ) IF identification of primary mice RGCs (40×). Scale bars : 50 µm. ( B ) Primary RGCs pretreated with PGB (50 µM, 100 µM, or 200 µM) for 24 hours were subjected to OGD/R. OGD/R induced a considerable decrease in cell viability, whereas PGB improved cell viability. The most significant improvement was observed at 100 µM. ( C , D ) Primary RGCs pretreated with MK2206 (5 µM) were treated with PGB (100 µM) for 24 hours prior to the onset of OGD/R. The ROS level was measured through the fluorescent probe, DCFH-DA, and the ratio of ROS + cells was measured. PGB pretreatment inhibited ROS generation, but MK2206 counteracted the effect. (40×). Scale bars : 50 µm. ( E ) Representative IF images of primary RGCs (40×). Scale bars : 50 µm. The ratios of RGCs expressing p-Akt, p-GSK3β, and β-catenin were downregulated after OGD/R, which was countered by PGB pretreatment. However, MK2206 decreased the ratios of positive cells ( n = 6). * P < 0.05 versus control group; ** P < 0.05 versus NaCl + OGD/R group; *** P < 0.05 versus PGB 100 µM + OGD/R group; **** P < 0.05 versus PGB + DMSO + OGD/R group.

    Techniques Used: Expressing

    PGB enhanced RGC survival following RIR injury via the Akt/GSK3β/β-catenin signaling pathway.
    Figure Legend Snippet: PGB enhanced RGC survival following RIR injury via the Akt/GSK3β/β-catenin signaling pathway.

    Techniques Used:

    phospho glycogen synthase kinase 3β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho glycogen synthase kinase 3β
    AMPK–ERK–PKC interplay in GBS PBMCs. ( a , b ) PBMCs were isolated from control subjects (CTRL; n = 20) and GBS patients ( n = 23), and the levels of phosphorylated ERK, p38 MAPK, and <t>GSK3β</t> were assessed by immunoblotting. The representative blots and densitometry data (relative to actin) are shown in ( a ), with horizontal lines representing median values and interquartile range (* p < 0.05, two-tailed Mann–Whitney U test). The correlations between GSK3β and phospho-AMPK, phospho-LKB1, or phospho-ERK are presented in ( b ) (r s —Spearman’s correlation coefficient; * p < 0.05, Spearman’s rank-order test).
    Phospho Glycogen Synthase Kinase 3β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Downregulation of LKB1/AMPK Signaling in Blood Mononuclear Cells Is Associated with the Severity of Guillain–Barre Syndrome"

    Article Title: Downregulation of LKB1/AMPK Signaling in Blood Mononuclear Cells Is Associated with the Severity of Guillain–Barre Syndrome

    Journal: Cells

    doi: 10.3390/cells11182897

    AMPK–ERK–PKC interplay in GBS PBMCs. ( a , b ) PBMCs were isolated from control subjects (CTRL; n = 20) and GBS patients ( n = 23), and the levels of phosphorylated ERK, p38 MAPK, and GSK3β were assessed by immunoblotting. The representative blots and densitometry data (relative to actin) are shown in ( a ), with horizontal lines representing median values and interquartile range (* p < 0.05, two-tailed Mann–Whitney U test). The correlations between GSK3β and phospho-AMPK, phospho-LKB1, or phospho-ERK are presented in ( b ) (r s —Spearman’s correlation coefficient; * p < 0.05, Spearman’s rank-order test).
    Figure Legend Snippet: AMPK–ERK–PKC interplay in GBS PBMCs. ( a , b ) PBMCs were isolated from control subjects (CTRL; n = 20) and GBS patients ( n = 23), and the levels of phosphorylated ERK, p38 MAPK, and GSK3β were assessed by immunoblotting. The representative blots and densitometry data (relative to actin) are shown in ( a ), with horizontal lines representing median values and interquartile range (* p < 0.05, two-tailed Mann–Whitney U test). The correlations between GSK3β and phospho-AMPK, phospho-LKB1, or phospho-ERK are presented in ( b ) (r s —Spearman’s correlation coefficient; * p < 0.05, Spearman’s rank-order test).

    Techniques Used: Isolation, Western Blot, Two Tailed Test, MANN-WHITNEY

    glycogen synthase kinase 3 beta  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc glycogen synthase kinase 3 beta
    Knockdown of PTS inhibited the Wnt pathway. (A,B) Transcriptome sequencing analysis after knockdown of PTS . (C,D) GSEA of PTS . (E,F) Protein levels of WNT5A , GSK-3β , β-catenin , and cyclin D1 were tested by Western blot. Cell viability, migration, and proliferation were detected using CCK-8 (G), wound healing assay (measured under a microscope, magnification 200×) (H), and transwell assay (stained with crystal violet; magnification 200×) (I). ## , P<0.05, compared with the si- PTS -1 group. *, P<0.05; **, P<0.01, compared with the si-NC group. sh-NC, shRNA-negative control; shRNA, short hairpin RNA; sh- PTS , shRNA for PTS ; PTS , 6-pyruvoyl-tetrahydropterin synthase; GSEA, Gene Set Enrichment Analysis; GSK-3β , glycogen synthase <t>kinase-3</t> beta; WNT5A , Wnt family member 5A; GAPDH , glyceraldehyde 3-phosphate dehydrogenase; si-NC, siRNA-negative control; siRNA, small interfering RNA; si- PTS , siRNA for PTS ; CCK-8, Cell Counting Kit-8.
    Glycogen Synthase Kinase 3 Beta, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "PTS is activated by ATF4 and promotes lung adenocarcinoma development via the Wnt pathway"

    Article Title: PTS is activated by ATF4 and promotes lung adenocarcinoma development via the Wnt pathway

    Journal: Translational Lung Cancer Research

    doi: 10.21037/tlcr-22-593

    Knockdown of PTS inhibited the Wnt pathway. (A,B) Transcriptome sequencing analysis after knockdown of PTS . (C,D) GSEA of PTS . (E,F) Protein levels of WNT5A , GSK-3β , β-catenin , and cyclin D1 were tested by Western blot. Cell viability, migration, and proliferation were detected using CCK-8 (G), wound healing assay (measured under a microscope, magnification 200×) (H), and transwell assay (stained with crystal violet; magnification 200×) (I). ## , P<0.05, compared with the si- PTS -1 group. *, P<0.05; **, P<0.01, compared with the si-NC group. sh-NC, shRNA-negative control; shRNA, short hairpin RNA; sh- PTS , shRNA for PTS ; PTS , 6-pyruvoyl-tetrahydropterin synthase; GSEA, Gene Set Enrichment Analysis; GSK-3β , glycogen synthase kinase-3 beta; WNT5A , Wnt family member 5A; GAPDH , glyceraldehyde 3-phosphate dehydrogenase; si-NC, siRNA-negative control; siRNA, small interfering RNA; si- PTS , siRNA for PTS ; CCK-8, Cell Counting Kit-8.
    Figure Legend Snippet: Knockdown of PTS inhibited the Wnt pathway. (A,B) Transcriptome sequencing analysis after knockdown of PTS . (C,D) GSEA of PTS . (E,F) Protein levels of WNT5A , GSK-3β , β-catenin , and cyclin D1 were tested by Western blot. Cell viability, migration, and proliferation were detected using CCK-8 (G), wound healing assay (measured under a microscope, magnification 200×) (H), and transwell assay (stained with crystal violet; magnification 200×) (I). ## , P<0.05, compared with the si- PTS -1 group. *, P<0.05; **, P<0.01, compared with the si-NC group. sh-NC, shRNA-negative control; shRNA, short hairpin RNA; sh- PTS , shRNA for PTS ; PTS , 6-pyruvoyl-tetrahydropterin synthase; GSEA, Gene Set Enrichment Analysis; GSK-3β , glycogen synthase kinase-3 beta; WNT5A , Wnt family member 5A; GAPDH , glyceraldehyde 3-phosphate dehydrogenase; si-NC, siRNA-negative control; siRNA, small interfering RNA; si- PTS , siRNA for PTS ; CCK-8, Cell Counting Kit-8.

    Techniques Used: Sequencing, Western Blot, Migration, CCK-8 Assay, Wound Healing Assay, Microscopy, Transwell Assay, Staining, shRNA, Negative Control, Small Interfering RNA, Cell Counting

    glycogen synthase kinase 3 α β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc glycogen synthase kinase 3 α β
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    p glycogen synthase kinase 3β  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti glycogen synthase kinase 3 beta
    Experimental scheme for this study. After gene identification at 1 month of age, 3-month-old male WT and 3×Tg-AD mice were randomly assigned to four groups with 10 animals each and then intragastrically administered either ICA or vehicle for 5 months (WT + vehicle, WT + ICA, 3×Tg-AD + vehicle, 3×Tg-AD + ICA groups). After performing behavior tests, the mice were euthanized. The cerebral cortexes were evaluated using HE and Nissl staining, immunofluorescent staining, and western blot assays to determine the above disease indicators. 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; Aβ: beta-amyloid protein; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; <t>GSK3β:</t> glycogen synthase <t>kinase</t> <t>3</t> beta; HE: hematoxylin and eosin; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; NeuN: neuronal nuclear antigen; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PSD95: postsynaptic density protein 95; WT: wild-type.
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    Molecular docking of core components and Wnt/β-catenin-related targets. Quercetin, luteolin, fisetin, wogonin, oroxylin A, boldine, tetrahydroalstonine, and galangin with (A) β-catenin, and (B) <t>GSK3β.</t> (C) Cluster heatmap of docking results. Numerical value indicated the binding energy ((kcal/mol).
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    Molecular docking of core components and Wnt/β-catenin-related targets. Quercetin, luteolin, fisetin, wogonin, oroxylin A, boldine, tetrahydroalstonine, and galangin with (A) β-catenin, and (B) <t>GSK3β.</t> (C) Cluster heatmap of docking results. Numerical value indicated the binding energy ((kcal/mol).
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    AMPK–ERK–PKC interplay in GBS PBMCs. ( a , b ) PBMCs were isolated from control subjects (CTRL; n = 20) and GBS patients ( n = 23), and the levels of phosphorylated ERK, p38 MAPK, and <t>GSK3β</t> were assessed by immunoblotting. The representative blots and densitometry data (relative to actin) are shown in ( a ), with horizontal lines representing median values and interquartile range (* p < 0.05, two-tailed Mann–Whitney U test). The correlations between GSK3β and phospho-AMPK, phospho-LKB1, or phospho-ERK are presented in ( b ) (r s —Spearman’s correlation coefficient; * p < 0.05, Spearman’s rank-order test).
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    Experimental scheme for this study. After gene identification at 1 month of age, 3-month-old male WT and 3×Tg-AD mice were randomly assigned to four groups with 10 animals each and then intragastrically administered either ICA or vehicle for 5 months (WT + vehicle, WT + ICA, 3×Tg-AD + vehicle, 3×Tg-AD + ICA groups). After performing behavior tests, the mice were euthanized. The cerebral cortexes were evaluated using HE and Nissl staining, immunofluorescent staining, and western blot assays to determine the above disease indicators. 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; Aβ: beta-amyloid protein; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; GSK3β: glycogen synthase kinase 3 beta; HE: hematoxylin and eosin; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; NeuN: neuronal nuclear antigen; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PSD95: postsynaptic density protein 95; WT: wild-type.

    Journal: Neural Regeneration Research

    Article Title: Icariin ameliorates memory deficits through regulating brain insulin signaling and glucose transporters in 3×Tg-AD mice

    doi: 10.4103/1673-5374.344840

    Figure Lengend Snippet: Experimental scheme for this study. After gene identification at 1 month of age, 3-month-old male WT and 3×Tg-AD mice were randomly assigned to four groups with 10 animals each and then intragastrically administered either ICA or vehicle for 5 months (WT + vehicle, WT + ICA, 3×Tg-AD + vehicle, 3×Tg-AD + ICA groups). After performing behavior tests, the mice were euthanized. The cerebral cortexes were evaluated using HE and Nissl staining, immunofluorescent staining, and western blot assays to determine the above disease indicators. 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; Aβ: beta-amyloid protein; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; GSK3β: glycogen synthase kinase 3 beta; HE: hematoxylin and eosin; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; NeuN: neuronal nuclear antigen; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PSD95: postsynaptic density protein 95; WT: wild-type.

    Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-insulin (1:1000; Proteintech, Cat# 15848-1-AP, RRID: AB_10597100), rabbit anti-insulin receptor substrate 1 (IRS1; 1:1000; Proteintech Cat# 17509-1-AP, RRID: AB_10596914), mouse anti-GLUT1 (1:1000; Proteintech, Cat# 66290-1-Ig, RRID: AB_2881673), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (1:50,000; Proteintech, Cat# 60004-1-Ig, RRID: AB_2107436), mouse anti-NeuN (1:1000, described above), rabbit anti-insulin receptor (IR) beta-subunit (1:1000; Cell Signaling Technology, Cat# 3025S, RRID: AB_2280448), rabbit anti-p-IRS1 Ser307 (1:1000; Cell Signaling Technology, Cat# 2381, RRID: AB_330342), rabbit anti-phosphatidylinositol 3-kinase (PI3K; 1:1000; Cell Signaling Technology, Cat# 4257, RRID: AB_659889), rabbit anti-phospho (p)-PI3K (1:1000; Cell Signaling Technology, Cat# 4228S, RRID: AB_659940), rabbit anti-protein kinase B (AKT; 1:1000; Cell Signaling Technology, Cat# 9272S, RRID: AB_329827), rabbit anti-glycogen synthase kinase 3 beta (GSK3β; 1:1000; Cell Signaling Technology, Cat# 9315, RRID: AB_490890), rabbit anti-p-GSK3β Ser9 (1:1000; Cell Signaling Technology, Cat# 9323, RRID: AB_2115201), rabbit anti-postsynaptic density protein 95 (PSD95; 1:1000; Abcam, Cat# ab18258, RRID: AB_444362), rabbit anti-APP (1:2000; Abcam, Cat# ab32136, RRID: AB_2289606), rabbit anti-Aβ 1–42 (1:1000; Abcam, Cat# ab201060, RRID: AB_2818982), rabbit anti-Aβ 1–40 (1:1000; Abcam, Cat# ab110888, RRID: AB_10890827), rabbit anti-PHF1 antibody (recognizing p-tau Ser396/404; 1:5000; Abcam, Cat# ab184951, RRID: AB_2861270), rabbit anti-p-tau Thr231 (1:5000; Abcam, Cat# ab151559, RRID: AB_2893278), rabbit anti-p-tau Ser199/202 (1:1000; Innovative Research, Cat# 44-768G, RRID: AB_1502103; Thermo Fisher Scientific, Waltham, MA, USA), rabbit anti-p-tau Thr217 (1:1000; Innovative Research, Cat# 44-744, RRID: AB_1502121), rabbit anti-p-IR Tyr1361 (1:1000; Thermo Fisher Scientific, Cat# PA5-38283, RRID: AB_2554884), rabbit anti-p-IRS1 Ser616 (1:1000, Innovative Research, Cat# 44-550G, RRID: AB_1501245), rabbit anti-p-AKT Ser473 (1:1000; Affinity Biosciences, Zhenjiang, China, Cat# AF0016, RRID: AB_2810275), and rabbit anti-GLUT3 (1:1000; Affinity Biosciences, Cat# AF5463, RRID: AB_2837947).

    Techniques: Staining, Western Blot, Transgenic Assay

    Effects of ICA on impaired insulin signaling in the cerebral cortex of 3×Tg-AD mice. (A) Insulin signaling: IR tyrosine autophosphorylation is stimulated by insulin and triggers IRS1 phosphorylation at tyrosine residues, which represents a positive regulatory mechanism that activates the PI3K/AKT pathway and results in the inhibition of GSK3β. However, serine phosphorylation of IRS1 at specific sites is a negative regulatory mechanism. (B) Representative expression patterns of molecules related to the insulin signaling pathway. (C) Quantification of proteins related to the insulin signaling pathway shown in (B). Protein levels were normalized to those in the WT + vehicle group. The data are presented as the means ± SEM ( n = 4–6). * P < 0.05, vs . WT + vehicle group; # P < 0.05, vs . 3×Tg-AD + vehicle group (one-way analysis of variance followed by the least significant difference test). 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; AKT: protein kinase B; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GSK3β: glycogen synthase kinase 3 beta; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PIP2: phosphatidylinositol (4,5) bisphosphate; PIP3: phosphatidylinositol (3,4,5) trisphosphate; PTEN: phosphatase and tensin homolog; WT: wild-type.

    Journal: Neural Regeneration Research

    Article Title: Icariin ameliorates memory deficits through regulating brain insulin signaling and glucose transporters in 3×Tg-AD mice

    doi: 10.4103/1673-5374.344840

    Figure Lengend Snippet: Effects of ICA on impaired insulin signaling in the cerebral cortex of 3×Tg-AD mice. (A) Insulin signaling: IR tyrosine autophosphorylation is stimulated by insulin and triggers IRS1 phosphorylation at tyrosine residues, which represents a positive regulatory mechanism that activates the PI3K/AKT pathway and results in the inhibition of GSK3β. However, serine phosphorylation of IRS1 at specific sites is a negative regulatory mechanism. (B) Representative expression patterns of molecules related to the insulin signaling pathway. (C) Quantification of proteins related to the insulin signaling pathway shown in (B). Protein levels were normalized to those in the WT + vehicle group. The data are presented as the means ± SEM ( n = 4–6). * P < 0.05, vs . WT + vehicle group; # P < 0.05, vs . 3×Tg-AD + vehicle group (one-way analysis of variance followed by the least significant difference test). 3×Tg-AD: A triple-transgenic mouse model of Alzheimer’s disease; AKT: protein kinase B; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GSK3β: glycogen synthase kinase 3 beta; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PIP2: phosphatidylinositol (4,5) bisphosphate; PIP3: phosphatidylinositol (3,4,5) trisphosphate; PTEN: phosphatase and tensin homolog; WT: wild-type.

    Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-insulin (1:1000; Proteintech, Cat# 15848-1-AP, RRID: AB_10597100), rabbit anti-insulin receptor substrate 1 (IRS1; 1:1000; Proteintech Cat# 17509-1-AP, RRID: AB_10596914), mouse anti-GLUT1 (1:1000; Proteintech, Cat# 66290-1-Ig, RRID: AB_2881673), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (1:50,000; Proteintech, Cat# 60004-1-Ig, RRID: AB_2107436), mouse anti-NeuN (1:1000, described above), rabbit anti-insulin receptor (IR) beta-subunit (1:1000; Cell Signaling Technology, Cat# 3025S, RRID: AB_2280448), rabbit anti-p-IRS1 Ser307 (1:1000; Cell Signaling Technology, Cat# 2381, RRID: AB_330342), rabbit anti-phosphatidylinositol 3-kinase (PI3K; 1:1000; Cell Signaling Technology, Cat# 4257, RRID: AB_659889), rabbit anti-phospho (p)-PI3K (1:1000; Cell Signaling Technology, Cat# 4228S, RRID: AB_659940), rabbit anti-protein kinase B (AKT; 1:1000; Cell Signaling Technology, Cat# 9272S, RRID: AB_329827), rabbit anti-glycogen synthase kinase 3 beta (GSK3β; 1:1000; Cell Signaling Technology, Cat# 9315, RRID: AB_490890), rabbit anti-p-GSK3β Ser9 (1:1000; Cell Signaling Technology, Cat# 9323, RRID: AB_2115201), rabbit anti-postsynaptic density protein 95 (PSD95; 1:1000; Abcam, Cat# ab18258, RRID: AB_444362), rabbit anti-APP (1:2000; Abcam, Cat# ab32136, RRID: AB_2289606), rabbit anti-Aβ 1–42 (1:1000; Abcam, Cat# ab201060, RRID: AB_2818982), rabbit anti-Aβ 1–40 (1:1000; Abcam, Cat# ab110888, RRID: AB_10890827), rabbit anti-PHF1 antibody (recognizing p-tau Ser396/404; 1:5000; Abcam, Cat# ab184951, RRID: AB_2861270), rabbit anti-p-tau Thr231 (1:5000; Abcam, Cat# ab151559, RRID: AB_2893278), rabbit anti-p-tau Ser199/202 (1:1000; Innovative Research, Cat# 44-768G, RRID: AB_1502103; Thermo Fisher Scientific, Waltham, MA, USA), rabbit anti-p-tau Thr217 (1:1000; Innovative Research, Cat# 44-744, RRID: AB_1502121), rabbit anti-p-IR Tyr1361 (1:1000; Thermo Fisher Scientific, Cat# PA5-38283, RRID: AB_2554884), rabbit anti-p-IRS1 Ser616 (1:1000, Innovative Research, Cat# 44-550G, RRID: AB_1501245), rabbit anti-p-AKT Ser473 (1:1000; Affinity Biosciences, Zhenjiang, China, Cat# AF0016, RRID: AB_2810275), and rabbit anti-GLUT3 (1:1000; Affinity Biosciences, Cat# AF5463, RRID: AB_2837947).

    Techniques: Inhibition, Expressing, Transgenic Assay

    Schematic diagram of the mechanism by which ICA regulates GLUTs and brain insulin signaling to ameliorate memory impairment in AD. Aβ: Amyloid-beta protein; AD: Alzheimer’s disease; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; GSK3β: glycogen synthase kinase 3 beta; G-tau: the attachment of O-linked N-acetylglucosamine (O-GlcNAc) on tau; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PIP2: phosphatidylinositol (4,5) bisphosphate; PIP3: phosphatidylinositol (3,4,5) trisphosphate; PTEN: phosphatase and tensin homolog.

    Journal: Neural Regeneration Research

    Article Title: Icariin ameliorates memory deficits through regulating brain insulin signaling and glucose transporters in 3×Tg-AD mice

    doi: 10.4103/1673-5374.344840

    Figure Lengend Snippet: Schematic diagram of the mechanism by which ICA regulates GLUTs and brain insulin signaling to ameliorate memory impairment in AD. Aβ: Amyloid-beta protein; AD: Alzheimer’s disease; AKT: protein kinase B; APP: amyloid precursor protein; GLUT: glucose transporter; GSK3β: glycogen synthase kinase 3 beta; G-tau: the attachment of O-linked N-acetylglucosamine (O-GlcNAc) on tau; ICA: icariin; IR: insulin receptor; IRS1: insulin receptor substrate 1; p: phosphorylation; PI3K: phosphatidylinositol 3-kinase; PIP2: phosphatidylinositol (4,5) bisphosphate; PIP3: phosphatidylinositol (3,4,5) trisphosphate; PTEN: phosphatase and tensin homolog.

    Article Snippet: The primary antibodies used in this study were as follows: rabbit anti-insulin (1:1000; Proteintech, Cat# 15848-1-AP, RRID: AB_10597100), rabbit anti-insulin receptor substrate 1 (IRS1; 1:1000; Proteintech Cat# 17509-1-AP, RRID: AB_10596914), mouse anti-GLUT1 (1:1000; Proteintech, Cat# 66290-1-Ig, RRID: AB_2881673), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (1:50,000; Proteintech, Cat# 60004-1-Ig, RRID: AB_2107436), mouse anti-NeuN (1:1000, described above), rabbit anti-insulin receptor (IR) beta-subunit (1:1000; Cell Signaling Technology, Cat# 3025S, RRID: AB_2280448), rabbit anti-p-IRS1 Ser307 (1:1000; Cell Signaling Technology, Cat# 2381, RRID: AB_330342), rabbit anti-phosphatidylinositol 3-kinase (PI3K; 1:1000; Cell Signaling Technology, Cat# 4257, RRID: AB_659889), rabbit anti-phospho (p)-PI3K (1:1000; Cell Signaling Technology, Cat# 4228S, RRID: AB_659940), rabbit anti-protein kinase B (AKT; 1:1000; Cell Signaling Technology, Cat# 9272S, RRID: AB_329827), rabbit anti-glycogen synthase kinase 3 beta (GSK3β; 1:1000; Cell Signaling Technology, Cat# 9315, RRID: AB_490890), rabbit anti-p-GSK3β Ser9 (1:1000; Cell Signaling Technology, Cat# 9323, RRID: AB_2115201), rabbit anti-postsynaptic density protein 95 (PSD95; 1:1000; Abcam, Cat# ab18258, RRID: AB_444362), rabbit anti-APP (1:2000; Abcam, Cat# ab32136, RRID: AB_2289606), rabbit anti-Aβ 1–42 (1:1000; Abcam, Cat# ab201060, RRID: AB_2818982), rabbit anti-Aβ 1–40 (1:1000; Abcam, Cat# ab110888, RRID: AB_10890827), rabbit anti-PHF1 antibody (recognizing p-tau Ser396/404; 1:5000; Abcam, Cat# ab184951, RRID: AB_2861270), rabbit anti-p-tau Thr231 (1:5000; Abcam, Cat# ab151559, RRID: AB_2893278), rabbit anti-p-tau Ser199/202 (1:1000; Innovative Research, Cat# 44-768G, RRID: AB_1502103; Thermo Fisher Scientific, Waltham, MA, USA), rabbit anti-p-tau Thr217 (1:1000; Innovative Research, Cat# 44-744, RRID: AB_1502121), rabbit anti-p-IR Tyr1361 (1:1000; Thermo Fisher Scientific, Cat# PA5-38283, RRID: AB_2554884), rabbit anti-p-IRS1 Ser616 (1:1000, Innovative Research, Cat# 44-550G, RRID: AB_1501245), rabbit anti-p-AKT Ser473 (1:1000; Affinity Biosciences, Zhenjiang, China, Cat# AF0016, RRID: AB_2810275), and rabbit anti-GLUT3 (1:1000; Affinity Biosciences, Cat# AF5463, RRID: AB_2837947).

    Techniques:

    Molecular docking of core components and Wnt/β-catenin-related targets. Quercetin, luteolin, fisetin, wogonin, oroxylin A, boldine, tetrahydroalstonine, and galangin with (A) β-catenin, and (B) GSK3β. (C) Cluster heatmap of docking results. Numerical value indicated the binding energy ((kcal/mol).

    Journal: Frontiers in Pharmacology

    Article Title: Stimulation of hair growth by Tianma Gouteng decoction: Identifying mechanisms based on chemical analysis, systems biology approach, and experimental evaluation

    doi: 10.3389/fphar.2022.1073392

    Figure Lengend Snippet: Molecular docking of core components and Wnt/β-catenin-related targets. Quercetin, luteolin, fisetin, wogonin, oroxylin A, boldine, tetrahydroalstonine, and galangin with (A) β-catenin, and (B) GSK3β. (C) Cluster heatmap of docking results. Numerical value indicated the binding energy ((kcal/mol).

    Article Snippet: Antibodies against β-catenin (8480S) and glycogen synthase kinase-3 beta (GSK3β; 12,456) were sourced from Cell Signaling Technology (Boston, United States).

    Techniques: Binding Assay

    TGD promoted hair shaft elongation in rat vibrissa follicles. (A) Photographs of rat vibrissa follicles cultured with TGD for 0, 3, 6, 9, 12 days; The double arrow area represents the measured hair shaft length. (B) Changes in length of hair shafts in the vibrissa follicles treated with TGD for 12 days. Data are presented as mean ± SEM ( n = 6). ** p < 0.01 compared with the control group. (C–E) The representative images of immunofluorescence staining in rat vibrissa follicles for β-catenin, GSK3β, LEF1 (×40), picture of magnification ×200 in white box.

    Journal: Frontiers in Pharmacology

    Article Title: Stimulation of hair growth by Tianma Gouteng decoction: Identifying mechanisms based on chemical analysis, systems biology approach, and experimental evaluation

    doi: 10.3389/fphar.2022.1073392

    Figure Lengend Snippet: TGD promoted hair shaft elongation in rat vibrissa follicles. (A) Photographs of rat vibrissa follicles cultured with TGD for 0, 3, 6, 9, 12 days; The double arrow area represents the measured hair shaft length. (B) Changes in length of hair shafts in the vibrissa follicles treated with TGD for 12 days. Data are presented as mean ± SEM ( n = 6). ** p < 0.01 compared with the control group. (C–E) The representative images of immunofluorescence staining in rat vibrissa follicles for β-catenin, GSK3β, LEF1 (×40), picture of magnification ×200 in white box.

    Article Snippet: Antibodies against β-catenin (8480S) and glycogen synthase kinase-3 beta (GSK3β; 12,456) were sourced from Cell Signaling Technology (Boston, United States).

    Techniques: Cell Culture, Immunofluorescence, Staining

    (A–E) The representative images and statistical graph of β-catenin, GSK3β, p-GSK3β and LEF1 in DPCs (×200), n = 3 per group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group. (F–H) Western blot analysis of Wnt/β-catenin signaling pathways related protein expression in DPCs, n = 4 per group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group.

    Journal: Frontiers in Pharmacology

    Article Title: Stimulation of hair growth by Tianma Gouteng decoction: Identifying mechanisms based on chemical analysis, systems biology approach, and experimental evaluation

    doi: 10.3389/fphar.2022.1073392

    Figure Lengend Snippet: (A–E) The representative images and statistical graph of β-catenin, GSK3β, p-GSK3β and LEF1 in DPCs (×200), n = 3 per group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group. (F–H) Western blot analysis of Wnt/β-catenin signaling pathways related protein expression in DPCs, n = 4 per group, * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. the control group.

    Article Snippet: Antibodies against β-catenin (8480S) and glycogen synthase kinase-3 beta (GSK3β; 12,456) were sourced from Cell Signaling Technology (Boston, United States).

    Techniques: Western Blot, Expressing

    (A) Immunoblot and densitometric analyses of (B) total and (C) phosphorylated glycogen synthase kinase 3 beta (GSK3β), p38 MAP Kinase (p38) and p44/42 MAPK (Erk) were performed in whole cell extracts from human fibroblasts deriving from control (CONT, white bars, N = 3), mutated parkin (PARK, dark grey bars, N = 3), mutated LRRK2 (LRRK2, light grey bars, N = 3) and idiopathic PD (PD, black bars, N = 3). For the quantitation, values of total protein were normalized on the level of GAPDH of the relative sample, whereas the levels of phosphorylated form were normalized on the values of total protein. All values are expressed as mean ± SEM. * p <0.05 and ** p <0.02 vs control, # p <0.05 vs PD according to ANOVA, Tukey HSD post hoc test.

    Journal: PLoS ONE

    Article Title: Microtubule Destabilization Is Shared by Genetic and Idiopathic Parkinson’s Disease Patient Fibroblasts

    doi: 10.1371/journal.pone.0037467

    Figure Lengend Snippet: (A) Immunoblot and densitometric analyses of (B) total and (C) phosphorylated glycogen synthase kinase 3 beta (GSK3β), p38 MAP Kinase (p38) and p44/42 MAPK (Erk) were performed in whole cell extracts from human fibroblasts deriving from control (CONT, white bars, N = 3), mutated parkin (PARK, dark grey bars, N = 3), mutated LRRK2 (LRRK2, light grey bars, N = 3) and idiopathic PD (PD, black bars, N = 3). For the quantitation, values of total protein were normalized on the level of GAPDH of the relative sample, whereas the levels of phosphorylated form were normalized on the values of total protein. All values are expressed as mean ± SEM. * p <0.05 and ** p <0.02 vs control, # p <0.05 vs PD according to ANOVA, Tukey HSD post hoc test.

    Article Snippet: Membranes were probed with the following antibodies: α-tubulin mouse IgG (clone B-5-1-2, Sigma-Aldrich, St Louis, MO); β-tubulin mouse IgG (clone Tub 2.1, Sigma-Aldrich); deTyr tubulin rabbit IgG (Chemicon, Temecula, CA); Tyr tubulin mouse IgG (clone TUB-1A2, Sigma-Aldrich); Ac tubulin mouse IgG (clone 6-11B-1, Sigma-Aldrich); microtubule-associated protein 1 light chain 3 rabbit IgG (Sigma-Aldrich); vimentin mouse IgG (clone V6, Sigma-Aldrich); actin mouse IgM (N350, Amersham, Little Chalfont, UK); Caspase 3 rabbit IgG (Enzo Life Sciences Ag., Lausen, Switzerland), GADPH mouse IgG (Biogenesis, Poole, UK); Heat Shock Protein 70 mouse IgG (clone 3A3, Chemicon); Glycogen synthase kinase 3 beta rabbit IgG (Abcam, Cambride, UK); Phospho-Glycogen synthase kinase 3 beta (Ser9) rabbit IgG (Cell Signaling Technology, Beverly, MA); p38 alpha MAP Kinase mouse IgG (clone L53F8, Cell Signaling Technology); Phospho-p38 MAP Kinase (Thr180/Tyr182) rabbit IgG (clone 3D7, Cell Signaling Technology); p44/42 MAPK (Erk1/2) rabbit IgG (clone 137F5, Cell Signaling Technology); Phospho-p44/42 MAPK (Thr202/Tyr204) rabbit IgG (clone D13.14.4E, Cell Signaling Technology); parkin mouse IgG (clone prk8, Sigma-Aldrich).

    Techniques: Western Blot, Quantitation Assay

    AMPK–ERK–PKC interplay in GBS PBMCs. ( a , b ) PBMCs were isolated from control subjects (CTRL; n = 20) and GBS patients ( n = 23), and the levels of phosphorylated ERK, p38 MAPK, and GSK3β were assessed by immunoblotting. The representative blots and densitometry data (relative to actin) are shown in ( a ), with horizontal lines representing median values and interquartile range (* p < 0.05, two-tailed Mann–Whitney U test). The correlations between GSK3β and phospho-AMPK, phospho-LKB1, or phospho-ERK are presented in ( b ) (r s —Spearman’s correlation coefficient; * p < 0.05, Spearman’s rank-order test).

    Journal: Cells

    Article Title: Downregulation of LKB1/AMPK Signaling in Blood Mononuclear Cells Is Associated with the Severity of Guillain–Barre Syndrome

    doi: 10.3390/cells11182897

    Figure Lengend Snippet: AMPK–ERK–PKC interplay in GBS PBMCs. ( a , b ) PBMCs were isolated from control subjects (CTRL; n = 20) and GBS patients ( n = 23), and the levels of phosphorylated ERK, p38 MAPK, and GSK3β were assessed by immunoblotting. The representative blots and densitometry data (relative to actin) are shown in ( a ), with horizontal lines representing median values and interquartile range (* p < 0.05, two-tailed Mann–Whitney U test). The correlations between GSK3β and phospho-AMPK, phospho-LKB1, or phospho-ERK are presented in ( b ) (r s —Spearman’s correlation coefficient; * p < 0.05, Spearman’s rank-order test).

    Article Snippet: After blocking with 10% milk powder, membranes were incubated with primary rabbit antibodies against SQSTM1 (NBP1-48320; Novus Biologicals, Littleton, CO, USA), NBR1 (#9891), LC3B (#2775), beclin-1 (#3495), ATG5 (#12994), LKB1 (#3047), phospho-LKB1 (Ser428; #3482), AMPKα (#2603), phospho-AMPKα (Thr172; #2535), Raptor (#2280), phospho-Raptor (Ser792; #2083), phospho-eukaryotic translation initiation factor 4E-binding protein 1 (4EBP1) (Thr37/46; #2855), phospho-AKT (Ser473; #9271), phospho-ERK1/2 (Thr202/Tyr204; #9101), phospho-p38 MAPK (Thr180/Tyr182; #9211), phospho-glycogen synthase kinase 3β (GSK3β, Ser9; #9322), and actin (#4968) as a loading control (all from Cell Signaling Technology, Cambridge, MA, USA).

    Techniques: Isolation, Western Blot, Two Tailed Test, MANN-WHITNEY