rabbit anti phosphorylated akt p akt  (Danaher Inc)


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    Danaher Inc rabbit anti phosphorylated akt p akt
    Expression of <t>phosphorylated-AKT</t> (p-AKT), AKT, phosphorylated-GSK3β (p-GSK3β), GSK3β, and β-catenin in the nucleus of the ipsilateral hemisphere (excluding the infarct area). (A) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. Protein expression levels were normalized to GAPDH (total protein) or H3 (nuclear protein) ( n = 3). Expression of nuclear β-catenin, p-AKT, and p-GSK3β were higher in rats of the TMS (+) group compared with the TMS (–) group at 3, 7, 14, and 28 days. (B) Immunofluorescence of nuclear β-catenin (red, Alexa Fluor® 555) and Nestin (green)-positive cells in the TMS (+), TMS (–), and Sham groups ( n = 5). Nuclear β-catenin was upregulated in the TMS (+) group (analysis of variance [ANOVA]). Scale bars: 25 μm. (C) Ki67 (red, Alexa Fluor® 555) and Nestin (green, Alexa Fluor® 488)-positive cells in the TMS (–), TMS (H), and TMS + LY groups ( n = 5). More Ki67 + /Nestin + cells were detected in the TMS (H) group. Scale bars: 25 μm. (D) CCK8 analysis. Cell viability was decreased in the TMS (H) + LY group ( n = 5). (E) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (A: Student's t -test; B, D, E: Brown-Forsythe and Welch ANOVA tests followed by Dunnett's T3 multiple comparison test). D3/7/14/28 TMS (–): rats with no TMS at 3/7/14/28 days group; D3/7/14/28 TMS (+): rats with 10 Hz TMS at 3/7/14/28 days group; GAPDH: glyceric acid phosphated hydrogenase; ns: no significant; NSC: neural stem cell; SHAM: sham-operation group; TMS: transcranial magnetic stimulation; TMS (–): no TMS group; TMS (+): high-frequency TMS with 10 Hz group; TMS(H): NSCs with 10 Hz high-frequency TMS group; TMS(H) + LY: NSCs with 10 Hz high-frequency TMS and LY294002 group.
    Rabbit Anti Phosphorylated Akt P Akt, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti phosphorylated akt p akt - by Bioz Stars, 2024-07
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    Images

    1) Product Images from "High-frequency repetitive transcranial magnetic stimulation promotes neural stem cell proliferation after ischemic stroke"

    Article Title: High-frequency repetitive transcranial magnetic stimulation promotes neural stem cell proliferation after ischemic stroke

    Journal: Neural Regeneration Research

    doi: 10.4103/1673-5374.389303

    Expression of phosphorylated-AKT (p-AKT), AKT, phosphorylated-GSK3β (p-GSK3β), GSK3β, and β-catenin in the nucleus of the ipsilateral hemisphere (excluding the infarct area). (A) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. Protein expression levels were normalized to GAPDH (total protein) or H3 (nuclear protein) ( n = 3). Expression of nuclear β-catenin, p-AKT, and p-GSK3β were higher in rats of the TMS (+) group compared with the TMS (–) group at 3, 7, 14, and 28 days. (B) Immunofluorescence of nuclear β-catenin (red, Alexa Fluor® 555) and Nestin (green)-positive cells in the TMS (+), TMS (–), and Sham groups ( n = 5). Nuclear β-catenin was upregulated in the TMS (+) group (analysis of variance [ANOVA]). Scale bars: 25 μm. (C) Ki67 (red, Alexa Fluor® 555) and Nestin (green, Alexa Fluor® 488)-positive cells in the TMS (–), TMS (H), and TMS + LY groups ( n = 5). More Ki67 + /Nestin + cells were detected in the TMS (H) group. Scale bars: 25 μm. (D) CCK8 analysis. Cell viability was decreased in the TMS (H) + LY group ( n = 5). (E) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (A: Student's t -test; B, D, E: Brown-Forsythe and Welch ANOVA tests followed by Dunnett's T3 multiple comparison test). D3/7/14/28 TMS (–): rats with no TMS at 3/7/14/28 days group; D3/7/14/28 TMS (+): rats with 10 Hz TMS at 3/7/14/28 days group; GAPDH: glyceric acid phosphated hydrogenase; ns: no significant; NSC: neural stem cell; SHAM: sham-operation group; TMS: transcranial magnetic stimulation; TMS (–): no TMS group; TMS (+): high-frequency TMS with 10 Hz group; TMS(H): NSCs with 10 Hz high-frequency TMS group; TMS(H) + LY: NSCs with 10 Hz high-frequency TMS and LY294002 group.
    Figure Legend Snippet: Expression of phosphorylated-AKT (p-AKT), AKT, phosphorylated-GSK3β (p-GSK3β), GSK3β, and β-catenin in the nucleus of the ipsilateral hemisphere (excluding the infarct area). (A) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. Protein expression levels were normalized to GAPDH (total protein) or H3 (nuclear protein) ( n = 3). Expression of nuclear β-catenin, p-AKT, and p-GSK3β were higher in rats of the TMS (+) group compared with the TMS (–) group at 3, 7, 14, and 28 days. (B) Immunofluorescence of nuclear β-catenin (red, Alexa Fluor® 555) and Nestin (green)-positive cells in the TMS (+), TMS (–), and Sham groups ( n = 5). Nuclear β-catenin was upregulated in the TMS (+) group (analysis of variance [ANOVA]). Scale bars: 25 μm. (C) Ki67 (red, Alexa Fluor® 555) and Nestin (green, Alexa Fluor® 488)-positive cells in the TMS (–), TMS (H), and TMS + LY groups ( n = 5). More Ki67 + /Nestin + cells were detected in the TMS (H) group. Scale bars: 25 μm. (D) CCK8 analysis. Cell viability was decreased in the TMS (H) + LY group ( n = 5). (E) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (A: Student's t -test; B, D, E: Brown-Forsythe and Welch ANOVA tests followed by Dunnett's T3 multiple comparison test). D3/7/14/28 TMS (–): rats with no TMS at 3/7/14/28 days group; D3/7/14/28 TMS (+): rats with 10 Hz TMS at 3/7/14/28 days group; GAPDH: glyceric acid phosphated hydrogenase; ns: no significant; NSC: neural stem cell; SHAM: sham-operation group; TMS: transcranial magnetic stimulation; TMS (–): no TMS group; TMS (+): high-frequency TMS with 10 Hz group; TMS(H): NSCs with 10 Hz high-frequency TMS group; TMS(H) + LY: NSCs with 10 Hz high-frequency TMS and LY294002 group.

    Techniques Used: Expressing, Western Blot, Immunofluorescence, Comparison

    rTMS activates AKT through the Ca 2+ /P2 calcium channel/CaM pathway. (A) Calcium imaging of NSCs by Fluo-4 AM labeling. Cells in the TMS (+) group showed stronger signal. Scale bars: 25 μm. (B, C) Levels of Ca 2+ influx by fluorescence intensity (B; n = 5) and flow cytometry (C; n = 4). (D) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Expression of nuclear β-catenin, p-GSK3β, and p-AKT were higher in NSCs of the TMS (H) group compared with other groups. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (B: Student's t -test; D: Brown-Forsythe and Welch analysis of variance tests followed by Dunnett's T3 multiple comparison test). 2-APB: 2-Aminoethyl diphenylborinate; CPZ: chlorpromazine; CTRL: NSCs with no OGD group; NSC: neural stem cell; OGD: NSCs with OGD group; p-AKT: phosphorylated-AKT; p-GSK3β: phosphorylated-glycogen synthase kinase-3β; TMS(H): NSCs with 10 Hz high-frequency TMS group; U7: inhibitor U73122.
    Figure Legend Snippet: rTMS activates AKT through the Ca 2+ /P2 calcium channel/CaM pathway. (A) Calcium imaging of NSCs by Fluo-4 AM labeling. Cells in the TMS (+) group showed stronger signal. Scale bars: 25 μm. (B, C) Levels of Ca 2+ influx by fluorescence intensity (B; n = 5) and flow cytometry (C; n = 4). (D) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Expression of nuclear β-catenin, p-GSK3β, and p-AKT were higher in NSCs of the TMS (H) group compared with other groups. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (B: Student's t -test; D: Brown-Forsythe and Welch analysis of variance tests followed by Dunnett's T3 multiple comparison test). 2-APB: 2-Aminoethyl diphenylborinate; CPZ: chlorpromazine; CTRL: NSCs with no OGD group; NSC: neural stem cell; OGD: NSCs with OGD group; p-AKT: phosphorylated-AKT; p-GSK3β: phosphorylated-glycogen synthase kinase-3β; TMS(H): NSCs with 10 Hz high-frequency TMS group; U7: inhibitor U73122.

    Techniques Used: Imaging, Labeling, Fluorescence, Flow Cytometry, Western Blot, Expressing, Comparison

    rabbit anti phosphorylated akt p akt  (Danaher Inc)


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    Danaher Inc rabbit anti phosphorylated akt p akt
    Rabbit Anti Phosphorylated Akt P Akt, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti phosphorylated akt p akt/product/Danaher Inc
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    phosphorylated p akt  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc phosphorylated p akt
    Recombinant adenovirus enhancing apoptosis through PI3K/AKT-caspase-3 ameliorates resistance to DDP on SKOV3 cells. (A) The representative images of the Annexin V-APC staining assay. (B) Recombinant adenovirus and/or DDP promoted the apoptosis of ovarian cancer cells and the apoptosis rate in combination group was significantly higher than that in DDP group (n=4 in each group). (C) PI3K was significantly suppressed by recombinant adenovirus and/or DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (D) AKT was significantly suppressed by recombinant adenovirus and (or) DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (E) Procaspase-3 was significantly reduced after recombinant adenovirus and/or DDP incubation, whereas cleaved caspase-3 was considerably increased (n=4 in each group). Data are shown as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 vs. corresponding controls; # P<0.05, ## P<0.01, ### P<0.001 vs. DDP groups, & P<0.05, &&& P<0.001 vs. DDP combined with Ad-NC groups. DDP, cisplatin; si, short interfering; NC, negative control; p-. <t>phosphorylated.</t>
    Phosphorylated P Akt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Dual‑regulated oncolytic adenovirus carrying ERCC1 ‑siRNA gene possesses potent antitumor effect on ovarian cancer cells"

    Article Title: Dual‑regulated oncolytic adenovirus carrying ERCC1 ‑siRNA gene possesses potent antitumor effect on ovarian cancer cells

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2024.13245

    Recombinant adenovirus enhancing apoptosis through PI3K/AKT-caspase-3 ameliorates resistance to DDP on SKOV3 cells. (A) The representative images of the Annexin V-APC staining assay. (B) Recombinant adenovirus and/or DDP promoted the apoptosis of ovarian cancer cells and the apoptosis rate in combination group was significantly higher than that in DDP group (n=4 in each group). (C) PI3K was significantly suppressed by recombinant adenovirus and/or DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (D) AKT was significantly suppressed by recombinant adenovirus and (or) DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (E) Procaspase-3 was significantly reduced after recombinant adenovirus and/or DDP incubation, whereas cleaved caspase-3 was considerably increased (n=4 in each group). Data are shown as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 vs. corresponding controls; # P<0.05, ## P<0.01, ### P<0.001 vs. DDP groups, & P<0.05, &&& P<0.001 vs. DDP combined with Ad-NC groups. DDP, cisplatin; si, short interfering; NC, negative control; p-. phosphorylated.
    Figure Legend Snippet: Recombinant adenovirus enhancing apoptosis through PI3K/AKT-caspase-3 ameliorates resistance to DDP on SKOV3 cells. (A) The representative images of the Annexin V-APC staining assay. (B) Recombinant adenovirus and/or DDP promoted the apoptosis of ovarian cancer cells and the apoptosis rate in combination group was significantly higher than that in DDP group (n=4 in each group). (C) PI3K was significantly suppressed by recombinant adenovirus and/or DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (D) AKT was significantly suppressed by recombinant adenovirus and (or) DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (E) Procaspase-3 was significantly reduced after recombinant adenovirus and/or DDP incubation, whereas cleaved caspase-3 was considerably increased (n=4 in each group). Data are shown as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 vs. corresponding controls; # P<0.05, ## P<0.01, ### P<0.001 vs. DDP groups, & P<0.05, &&& P<0.001 vs. DDP combined with Ad-NC groups. DDP, cisplatin; si, short interfering; NC, negative control; p-. phosphorylated.

    Techniques Used: Recombinant, Staining, Incubation, Negative Control

    phosphorylated p akt  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc phosphorylated p akt
    Effects of madanin-1 WT and madanin-1 2S on signaling pathways. SKOV3 and MDA-MB-231 cells were pretreated with madanin-1 WT or madanin-1 2S (10 µg/ml for 30 min) prior to treatment with thrombin (2 U/ml for 15 min). The protein expression levels of <t>p-Akt,</t> t-Akt, p-ERK and t-ERK in SKOV3 and MDA-MB-231 cells were analyzed by western blotting. (A) Representative gel images. (B) In SKOV3 cells, thrombin-induced p-Akt and p-ERK expression was significantly attenuated by both madanin-1 WT and madanin-1 2S treatment. In MDA-MB-231 cells, thrombin-induced p-Akt and p-ERK expression was significantly inhibited only by madanin-1 2S treatment. The protein expression levels were normalized to the level of β-actin in the same sample. Data are presented as the mean relative density ratio of each protein <t>(phosphorylated</t> form/total form) ± standard deviation of three independent experiments completed in triplicate (*P<0.05). WT, wild-type; 2S, 2 sulfation; p, phosphorylated; t, total; ERK, extracellular signal-regulated kinase.
    Phosphorylated P Akt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Inhibitory effect of recombinant tyrosine‑sulfated madanin‑1, a thrombin inhibitor, on the behavior of MDA‑MB‑231 and SKOV3 cells in vitro"

    Article Title: Inhibitory effect of recombinant tyrosine‑sulfated madanin‑1, a thrombin inhibitor, on the behavior of MDA‑MB‑231 and SKOV3 cells in vitro

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2024.13238

    Effects of madanin-1 WT and madanin-1 2S on signaling pathways. SKOV3 and MDA-MB-231 cells were pretreated with madanin-1 WT or madanin-1 2S (10 µg/ml for 30 min) prior to treatment with thrombin (2 U/ml for 15 min). The protein expression levels of p-Akt, t-Akt, p-ERK and t-ERK in SKOV3 and MDA-MB-231 cells were analyzed by western blotting. (A) Representative gel images. (B) In SKOV3 cells, thrombin-induced p-Akt and p-ERK expression was significantly attenuated by both madanin-1 WT and madanin-1 2S treatment. In MDA-MB-231 cells, thrombin-induced p-Akt and p-ERK expression was significantly inhibited only by madanin-1 2S treatment. The protein expression levels were normalized to the level of β-actin in the same sample. Data are presented as the mean relative density ratio of each protein (phosphorylated form/total form) ± standard deviation of three independent experiments completed in triplicate (*P<0.05). WT, wild-type; 2S, 2 sulfation; p, phosphorylated; t, total; ERK, extracellular signal-regulated kinase.
    Figure Legend Snippet: Effects of madanin-1 WT and madanin-1 2S on signaling pathways. SKOV3 and MDA-MB-231 cells were pretreated with madanin-1 WT or madanin-1 2S (10 µg/ml for 30 min) prior to treatment with thrombin (2 U/ml for 15 min). The protein expression levels of p-Akt, t-Akt, p-ERK and t-ERK in SKOV3 and MDA-MB-231 cells were analyzed by western blotting. (A) Representative gel images. (B) In SKOV3 cells, thrombin-induced p-Akt and p-ERK expression was significantly attenuated by both madanin-1 WT and madanin-1 2S treatment. In MDA-MB-231 cells, thrombin-induced p-Akt and p-ERK expression was significantly inhibited only by madanin-1 2S treatment. The protein expression levels were normalized to the level of β-actin in the same sample. Data are presented as the mean relative density ratio of each protein (phosphorylated form/total form) ± standard deviation of three independent experiments completed in triplicate (*P<0.05). WT, wild-type; 2S, 2 sulfation; p, phosphorylated; t, total; ERK, extracellular signal-regulated kinase.

    Techniques Used: Expressing, Western Blot, Standard Deviation


    Structured Review

    Santa Cruz Biotechnology phosphorylated akt p akt
    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of <t>phosphorylated-PI3K</t> (p-PI3K), PI3K, <t>phosphorylated-Akt</t> <t>(p-Akt),</t> and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Phosphorylated Akt P Akt, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    1) Product Images from "Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes"

    Article Title: Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes

    Journal: Preventive Nutrition and Food Science

    doi: 10.3746/pnf.2024.29.2.154

    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-PI3K (p-PI3K), PI3K, phosphorylated-Akt (p-Akt), and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Figure Legend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-PI3K (p-PI3K), PI3K, phosphorylated-Akt (p-Akt), and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Techniques Used: Western Blot, Expressing, Control

    Effects of Agastache rugosa extract (ARE) and tilianin on the mammalian target of rapamycin (mTOR) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-mTOR (p-mTOR), mTOR, phosphorylated-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phosphorylated-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), and 4EBP1 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Figure Legend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on the mammalian target of rapamycin (mTOR) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-mTOR (p-mTOR), mTOR, phosphorylated-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phosphorylated-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), and 4EBP1 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Techniques Used: Western Blot, Expressing, Binding Assay, Control

    Effects of Agastache rugosa extract (ARE) and tilianin on protein-degradation-related pathways in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-forkhead box class O 3 (p-FoxO3) and FoxO3 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Reverse transcription polymerase chain reaction was used to determine the mRNA expression levels of muscle RING-finger protein-1 (MuRF1) and atrogin-1 in response to (C) ARE and (D) tilianin. β-Actin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. # P <0.05, ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Figure Legend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on protein-degradation-related pathways in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-forkhead box class O 3 (p-FoxO3) and FoxO3 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Reverse transcription polymerase chain reaction was used to determine the mRNA expression levels of muscle RING-finger protein-1 (MuRF1) and atrogin-1 in response to (C) ARE and (D) tilianin. β-Actin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. # P <0.05, ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Techniques Used: Western Blot, Expressing, Control, Reverse Transcription, Polymerase Chain Reaction

    phosphorylated akt p akt  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc phosphorylated akt p akt
    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of <t>phosphorylated-PI3K</t> (p-PI3K), PI3K, <t>phosphorylated-Akt</t> <t>(p-Akt),</t> and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Phosphorylated Akt P Akt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated akt p akt/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    phosphorylated akt p akt - by Bioz Stars, 2024-07
    86/100 stars

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    1) Product Images from "Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes"

    Article Title: Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes

    Journal: Preventive Nutrition and Food Science

    doi: 10.3746/pnf.2024.29.2.154

    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-PI3K (p-PI3K), PI3K, phosphorylated-Akt (p-Akt), and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Figure Legend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-PI3K (p-PI3K), PI3K, phosphorylated-Akt (p-Akt), and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Techniques Used: Western Blot, Expressing, Control

    Effects of Agastache rugosa extract (ARE) and tilianin on the mammalian target of rapamycin (mTOR) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-mTOR (p-mTOR), mTOR, phosphorylated-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phosphorylated-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), and 4EBP1 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Figure Legend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on the mammalian target of rapamycin (mTOR) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-mTOR (p-mTOR), mTOR, phosphorylated-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phosphorylated-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), and 4EBP1 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Techniques Used: Western Blot, Expressing, Binding Assay, Control

    Effects of Agastache rugosa extract (ARE) and tilianin on protein-degradation-related pathways in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-forkhead box class O 3 (p-FoxO3) and FoxO3 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Reverse transcription polymerase chain reaction was used to determine the mRNA expression levels of muscle RING-finger protein-1 (MuRF1) and atrogin-1 in response to (C) ARE and (D) tilianin. β-Actin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. # P <0.05, ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Figure Legend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on protein-degradation-related pathways in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-forkhead box class O 3 (p-FoxO3) and FoxO3 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Reverse transcription polymerase chain reaction was used to determine the mRNA expression levels of muscle RING-finger protein-1 (MuRF1) and atrogin-1 in response to (C) ARE and (D) tilianin. β-Actin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. # P <0.05, ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Techniques Used: Western Blot, Expressing, Control, Reverse Transcription, Polymerase Chain Reaction


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    Affinity Biosciences anti phosphorylated akt p akt
    Anti Phosphorylated Akt P Akt, supplied by Affinity Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phosphorylated akt p akt/product/Affinity Biosciences
    Average 86 stars, based on 1 article reviews
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    anti phosphorylated akt p akt  (Danaher Inc)


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    Danaher Inc anti phosphorylated akt p akt
    Anti Phosphorylated Akt P Akt, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phosphorylated akt p akt/product/Danaher Inc
    Average 86 stars, based on 1 article reviews
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    phosphorylated akt p akt  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phosphorylated akt p akt
    ARPE-19 cells were stimulated by 100 ng/mL of EGF for 24h. A: The images of cells morphology were obtained after treatment, scale bar: 100 µm. B: Cellular viability was tested by the MTT assay, n=6. C, D: Cellular proliferation was analyzed by the BrdU labeling assay, scale bar: 50 µm, n=3. E, F: Cellular migration was measured by the wound healing assay, scale bar: 200 µm, n=3. G, H: After treatment with EGF (100 ng/mL) for 0, 0.25, 0.5, 1.0, 3.0, and 6.0h, EGFR, p-EGFR, <t>AKT</t> and <t>p-AKT</t> proteins in RPE cells were detected by Western blot assay. aP<0.05, bP<0.01. EGF: Epidermal growth factor; APRE-19: Adult retinalpigment epithelial cell line-19; EGFR: Epithelial growth factor receptor; AKT: Protein kinase B; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.
    Phosphorylated Akt P Akt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated akt p akt/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    phosphorylated akt p akt - by Bioz Stars, 2024-07
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    1) Product Images from "Inhibition of EGFR attenuates EGF-induced activation of retinal pigment epithelium cell via EGFR/AKT signaling pathway"

    Article Title: Inhibition of EGFR attenuates EGF-induced activation of retinal pigment epithelium cell via EGFR/AKT signaling pathway

    Journal: International Journal of Ophthalmology

    doi: 10.18240/ijo.2024.06.05

    ARPE-19 cells were stimulated by 100 ng/mL of EGF for 24h. A: The images of cells morphology were obtained after treatment, scale bar: 100 µm. B: Cellular viability was tested by the MTT assay, n=6. C, D: Cellular proliferation was analyzed by the BrdU labeling assay, scale bar: 50 µm, n=3. E, F: Cellular migration was measured by the wound healing assay, scale bar: 200 µm, n=3. G, H: After treatment with EGF (100 ng/mL) for 0, 0.25, 0.5, 1.0, 3.0, and 6.0h, EGFR, p-EGFR, AKT and p-AKT proteins in RPE cells were detected by Western blot assay. aP<0.05, bP<0.01. EGF: Epidermal growth factor; APRE-19: Adult retinalpigment epithelial cell line-19; EGFR: Epithelial growth factor receptor; AKT: Protein kinase B; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.
    Figure Legend Snippet: ARPE-19 cells were stimulated by 100 ng/mL of EGF for 24h. A: The images of cells morphology were obtained after treatment, scale bar: 100 µm. B: Cellular viability was tested by the MTT assay, n=6. C, D: Cellular proliferation was analyzed by the BrdU labeling assay, scale bar: 50 µm, n=3. E, F: Cellular migration was measured by the wound healing assay, scale bar: 200 µm, n=3. G, H: After treatment with EGF (100 ng/mL) for 0, 0.25, 0.5, 1.0, 3.0, and 6.0h, EGFR, p-EGFR, AKT and p-AKT proteins in RPE cells were detected by Western blot assay. aP<0.05, bP<0.01. EGF: Epidermal growth factor; APRE-19: Adult retinalpigment epithelial cell line-19; EGFR: Epithelial growth factor receptor; AKT: Protein kinase B; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.

    Techniques Used: MTT Assay, Labeling, Migration, Wound Healing Assay, Western Blot

    A, B: ARPE-19 cells were treated with erlotinib (0, 5, 10, 20, and 50 µmol/L) for 24h. The expreesion of total EGFR and AKT proteins were measured by Western blot. aP<0.05, bP<0.01. C: After treatment with 50 µmol/L erlotinib for 24h, EGFR, F-actin, and nucleus in ARPE-19 cells were measured by immunofluorescence staining, scale bar: 50 µm. APRE-19: Adult retinal pigment epithelial cell line-19; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; DAPI: Diamidino-2-phenylindole.
    Figure Legend Snippet: A, B: ARPE-19 cells were treated with erlotinib (0, 5, 10, 20, and 50 µmol/L) for 24h. The expreesion of total EGFR and AKT proteins were measured by Western blot. aP<0.05, bP<0.01. C: After treatment with 50 µmol/L erlotinib for 24h, EGFR, F-actin, and nucleus in ARPE-19 cells were measured by immunofluorescence staining, scale bar: 50 µm. APRE-19: Adult retinal pigment epithelial cell line-19; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; DAPI: Diamidino-2-phenylindole.

    Techniques Used: Western Blot, Immunofluorescence, Staining

    ARPE-19 cells were pretreated by erlotinib (20 µmol/L) for 12h and then stimulated with 100 ng/mL of EGF for 24h. A, B: BrdU staining assay was used to test cellular proliferation, scale bar: 50 µm, n=3. C: Cellular viability was sized by MTT assay, n=8. D, E: Cellular migration was tested by the wound healing assay, scale bar: 200 µm, n=3. F, G: ARPE-19 cells were pretreated with 20 µmol/L erlotinib for 12h and then treated with EGF (100 ng/mL) for 15min. Western blotting was used to detected EGFR/AKT signaling pathway proteins expressions. H, I: After pretreatment with 20 µmol/L erlotinib for 12h, ARPE-19 cells were treated with EGF (100 ng/mL) for 24h. N-cadherin, α-SMA and vimentin proteins were determined by Western blot. aP<0.05, bP<0.01. EGF: Epidermal growth factor; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; APRE-19: Adult retinal pigment epithelial cell line-19; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; α-SMA: α-smooth muscle actin; DAPI: Diamidino-2-phenylindole.
    Figure Legend Snippet: ARPE-19 cells were pretreated by erlotinib (20 µmol/L) for 12h and then stimulated with 100 ng/mL of EGF for 24h. A, B: BrdU staining assay was used to test cellular proliferation, scale bar: 50 µm, n=3. C: Cellular viability was sized by MTT assay, n=8. D, E: Cellular migration was tested by the wound healing assay, scale bar: 200 µm, n=3. F, G: ARPE-19 cells were pretreated with 20 µmol/L erlotinib for 12h and then treated with EGF (100 ng/mL) for 15min. Western blotting was used to detected EGFR/AKT signaling pathway proteins expressions. H, I: After pretreatment with 20 µmol/L erlotinib for 12h, ARPE-19 cells were treated with EGF (100 ng/mL) for 24h. N-cadherin, α-SMA and vimentin proteins were determined by Western blot. aP<0.05, bP<0.01. EGF: Epidermal growth factor; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; APRE-19: Adult retinal pigment epithelial cell line-19; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; α-SMA: α-smooth muscle actin; DAPI: Diamidino-2-phenylindole.

    Techniques Used: BrdU Staining, MTT Assay, Migration, Wound Healing Assay, Western Blot

    A, B: After EGFR-siRNA or control-siRNA were transfected into ARPE-19 cells, EGFR and AKT proteins were tested and analyzed by Western blotting. After EGFR-siRNA transfection, ARPE-19 cells were treated without or with 100 ng/mL EGF for 24h. C: Cellular viability was tested by MTT assay, n=8. D, E: Cellular proliferation was tested by the BrdU labeling assay, scale bar: 50 µm, n=3. F, G: Cellular migration was detected by the wound healing assay, scale bar: 200 µm, n=3. H, I: After transfected by control-siRNA or EGFR-siRNA, ARPE-19 cells were stimulated without or with 100 ng/mL of EGF for 15min. EGFR, p-EGFR, AKT and p-AKT protein were detected by Western blot. aP<0.05, bP<0.01. siRNA: Small interfering RNA; EGF: Epidermal growth factor; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; APRE-19: Adult retinal pigment epithelial cell line-19; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.
    Figure Legend Snippet: A, B: After EGFR-siRNA or control-siRNA were transfected into ARPE-19 cells, EGFR and AKT proteins were tested and analyzed by Western blotting. After EGFR-siRNA transfection, ARPE-19 cells were treated without or with 100 ng/mL EGF for 24h. C: Cellular viability was tested by MTT assay, n=8. D, E: Cellular proliferation was tested by the BrdU labeling assay, scale bar: 50 µm, n=3. F, G: Cellular migration was detected by the wound healing assay, scale bar: 200 µm, n=3. H, I: After transfected by control-siRNA or EGFR-siRNA, ARPE-19 cells were stimulated without or with 100 ng/mL of EGF for 15min. EGFR, p-EGFR, AKT and p-AKT protein were detected by Western blot. aP<0.05, bP<0.01. siRNA: Small interfering RNA; EGF: Epidermal growth factor; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; APRE-19: Adult retinal pigment epithelial cell line-19; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.

    Techniques Used: Control, Transfection, Western Blot, MTT Assay, Labeling, Migration, Wound Healing Assay, Small Interfering RNA

    phosphorylated akt p akt  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phosphorylated akt p akt
    ARPE-19 cells were stimulated by 100 ng/mL of EGF for 24h. A: The images of cells morphology were obtained after treatment, scale bar: 100 µm. B: Cellular viability was tested by the MTT assay, n=6. C, D: Cellular proliferation was analyzed by the BrdU labeling assay, scale bar: 50 µm, n=3. E, F: Cellular migration was measured by the wound healing assay, scale bar: 200 µm, n=3. G, H: After treatment with EGF (100 ng/mL) for 0, 0.25, 0.5, 1.0, 3.0, and 6.0h, EGFR, p-EGFR, <t>AKT</t> and <t>p-AKT</t> proteins in RPE cells were detected by Western blot assay. aP<0.05, bP<0.01. EGF: Epidermal growth factor; APRE-19: Adult retinalpigment epithelial cell line-19; EGFR: Epithelial growth factor receptor; AKT: Protein kinase B; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.
    Phosphorylated Akt P Akt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated akt p akt/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphorylated akt p akt - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Inhibition of EGFR attenuates EGF-induced activation of retinal pigment epithelium cell via EGFR/AKT signaling pathway"

    Article Title: Inhibition of EGFR attenuates EGF-induced activation of retinal pigment epithelium cell via EGFR/AKT signaling pathway

    Journal: International Journal of Ophthalmology

    doi: 10.18240/ijo.2024.06.05

    ARPE-19 cells were stimulated by 100 ng/mL of EGF for 24h. A: The images of cells morphology were obtained after treatment, scale bar: 100 µm. B: Cellular viability was tested by the MTT assay, n=6. C, D: Cellular proliferation was analyzed by the BrdU labeling assay, scale bar: 50 µm, n=3. E, F: Cellular migration was measured by the wound healing assay, scale bar: 200 µm, n=3. G, H: After treatment with EGF (100 ng/mL) for 0, 0.25, 0.5, 1.0, 3.0, and 6.0h, EGFR, p-EGFR, AKT and p-AKT proteins in RPE cells were detected by Western blot assay. aP<0.05, bP<0.01. EGF: Epidermal growth factor; APRE-19: Adult retinalpigment epithelial cell line-19; EGFR: Epithelial growth factor receptor; AKT: Protein kinase B; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.
    Figure Legend Snippet: ARPE-19 cells were stimulated by 100 ng/mL of EGF for 24h. A: The images of cells morphology were obtained after treatment, scale bar: 100 µm. B: Cellular viability was tested by the MTT assay, n=6. C, D: Cellular proliferation was analyzed by the BrdU labeling assay, scale bar: 50 µm, n=3. E, F: Cellular migration was measured by the wound healing assay, scale bar: 200 µm, n=3. G, H: After treatment with EGF (100 ng/mL) for 0, 0.25, 0.5, 1.0, 3.0, and 6.0h, EGFR, p-EGFR, AKT and p-AKT proteins in RPE cells were detected by Western blot assay. aP<0.05, bP<0.01. EGF: Epidermal growth factor; APRE-19: Adult retinalpigment epithelial cell line-19; EGFR: Epithelial growth factor receptor; AKT: Protein kinase B; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.

    Techniques Used: MTT Assay, Labeling, Migration, Wound Healing Assay, Western Blot

    A, B: ARPE-19 cells were treated with erlotinib (0, 5, 10, 20, and 50 µmol/L) for 24h. The expreesion of total EGFR and AKT proteins were measured by Western blot. aP<0.05, bP<0.01. C: After treatment with 50 µmol/L erlotinib for 24h, EGFR, F-actin, and nucleus in ARPE-19 cells were measured by immunofluorescence staining, scale bar: 50 µm. APRE-19: Adult retinal pigment epithelial cell line-19; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; DAPI: Diamidino-2-phenylindole.
    Figure Legend Snippet: A, B: ARPE-19 cells were treated with erlotinib (0, 5, 10, 20, and 50 µmol/L) for 24h. The expreesion of total EGFR and AKT proteins were measured by Western blot. aP<0.05, bP<0.01. C: After treatment with 50 µmol/L erlotinib for 24h, EGFR, F-actin, and nucleus in ARPE-19 cells were measured by immunofluorescence staining, scale bar: 50 µm. APRE-19: Adult retinal pigment epithelial cell line-19; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; DAPI: Diamidino-2-phenylindole.

    Techniques Used: Western Blot, Immunofluorescence, Staining

    ARPE-19 cells were pretreated by erlotinib (20 µmol/L) for 12h and then stimulated with 100 ng/mL of EGF for 24h. A, B: BrdU staining assay was used to test cellular proliferation, scale bar: 50 µm, n=3. C: Cellular viability was sized by MTT assay, n=8. D, E: Cellular migration was tested by the wound healing assay, scale bar: 200 µm, n=3. F, G: ARPE-19 cells were pretreated with 20 µmol/L erlotinib for 12h and then treated with EGF (100 ng/mL) for 15min. Western blotting was used to detected EGFR/AKT signaling pathway proteins expressions. H, I: After pretreatment with 20 µmol/L erlotinib for 12h, ARPE-19 cells were treated with EGF (100 ng/mL) for 24h. N-cadherin, α-SMA and vimentin proteins were determined by Western blot. aP<0.05, bP<0.01. EGF: Epidermal growth factor; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; APRE-19: Adult retinal pigment epithelial cell line-19; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; α-SMA: α-smooth muscle actin; DAPI: Diamidino-2-phenylindole.
    Figure Legend Snippet: ARPE-19 cells were pretreated by erlotinib (20 µmol/L) for 12h and then stimulated with 100 ng/mL of EGF for 24h. A, B: BrdU staining assay was used to test cellular proliferation, scale bar: 50 µm, n=3. C: Cellular viability was sized by MTT assay, n=8. D, E: Cellular migration was tested by the wound healing assay, scale bar: 200 µm, n=3. F, G: ARPE-19 cells were pretreated with 20 µmol/L erlotinib for 12h and then treated with EGF (100 ng/mL) for 15min. Western blotting was used to detected EGFR/AKT signaling pathway proteins expressions. H, I: After pretreatment with 20 µmol/L erlotinib for 12h, ARPE-19 cells were treated with EGF (100 ng/mL) for 24h. N-cadherin, α-SMA and vimentin proteins were determined by Western blot. aP<0.05, bP<0.01. EGF: Epidermal growth factor; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; APRE-19: Adult retinal pigment epithelial cell line-19; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; α-SMA: α-smooth muscle actin; DAPI: Diamidino-2-phenylindole.

    Techniques Used: BrdU Staining, MTT Assay, Migration, Wound Healing Assay, Western Blot

    A, B: After EGFR-siRNA or control-siRNA were transfected into ARPE-19 cells, EGFR and AKT proteins were tested and analyzed by Western blotting. After EGFR-siRNA transfection, ARPE-19 cells were treated without or with 100 ng/mL EGF for 24h. C: Cellular viability was tested by MTT assay, n=8. D, E: Cellular proliferation was tested by the BrdU labeling assay, scale bar: 50 µm, n=3. F, G: Cellular migration was detected by the wound healing assay, scale bar: 200 µm, n=3. H, I: After transfected by control-siRNA or EGFR-siRNA, ARPE-19 cells were stimulated without or with 100 ng/mL of EGF for 15min. EGFR, p-EGFR, AKT and p-AKT protein were detected by Western blot. aP<0.05, bP<0.01. siRNA: Small interfering RNA; EGF: Epidermal growth factor; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; APRE-19: Adult retinal pigment epithelial cell line-19; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.
    Figure Legend Snippet: A, B: After EGFR-siRNA or control-siRNA were transfected into ARPE-19 cells, EGFR and AKT proteins were tested and analyzed by Western blotting. After EGFR-siRNA transfection, ARPE-19 cells were treated without or with 100 ng/mL EGF for 24h. C: Cellular viability was tested by MTT assay, n=8. D, E: Cellular proliferation was tested by the BrdU labeling assay, scale bar: 50 µm, n=3. F, G: Cellular migration was detected by the wound healing assay, scale bar: 200 µm, n=3. H, I: After transfected by control-siRNA or EGFR-siRNA, ARPE-19 cells were stimulated without or with 100 ng/mL of EGF for 15min. EGFR, p-EGFR, AKT and p-AKT protein were detected by Western blot. aP<0.05, bP<0.01. siRNA: Small interfering RNA; EGF: Epidermal growth factor; EGFR: Epidermal growth factor receptor; AKT: Protein kinase B; APRE-19: Adult retinal pigment epithelial cell line-19; MTT: Methyl thiazolyl tetrazolium; BrdU: 5-Bromodeoxyuridinc; DAPI: Diamidino-2-phenylindole.

    Techniques Used: Control, Transfection, Western Blot, MTT Assay, Labeling, Migration, Wound Healing Assay, Small Interfering RNA

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    Danaher Inc rabbit anti phosphorylated akt p akt
    Expression of <t>phosphorylated-AKT</t> (p-AKT), AKT, phosphorylated-GSK3β (p-GSK3β), GSK3β, and β-catenin in the nucleus of the ipsilateral hemisphere (excluding the infarct area). (A) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. Protein expression levels were normalized to GAPDH (total protein) or H3 (nuclear protein) ( n = 3). Expression of nuclear β-catenin, p-AKT, and p-GSK3β were higher in rats of the TMS (+) group compared with the TMS (–) group at 3, 7, 14, and 28 days. (B) Immunofluorescence of nuclear β-catenin (red, Alexa Fluor® 555) and Nestin (green)-positive cells in the TMS (+), TMS (–), and Sham groups ( n = 5). Nuclear β-catenin was upregulated in the TMS (+) group (analysis of variance [ANOVA]). Scale bars: 25 μm. (C) Ki67 (red, Alexa Fluor® 555) and Nestin (green, Alexa Fluor® 488)-positive cells in the TMS (–), TMS (H), and TMS + LY groups ( n = 5). More Ki67 + /Nestin + cells were detected in the TMS (H) group. Scale bars: 25 μm. (D) CCK8 analysis. Cell viability was decreased in the TMS (H) + LY group ( n = 5). (E) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (A: Student's t -test; B, D, E: Brown-Forsythe and Welch ANOVA tests followed by Dunnett's T3 multiple comparison test). D3/7/14/28 TMS (–): rats with no TMS at 3/7/14/28 days group; D3/7/14/28 TMS (+): rats with 10 Hz TMS at 3/7/14/28 days group; GAPDH: glyceric acid phosphated hydrogenase; ns: no significant; NSC: neural stem cell; SHAM: sham-operation group; TMS: transcranial magnetic stimulation; TMS (–): no TMS group; TMS (+): high-frequency TMS with 10 Hz group; TMS(H): NSCs with 10 Hz high-frequency TMS group; TMS(H) + LY: NSCs with 10 Hz high-frequency TMS and LY294002 group.
    Rabbit Anti Phosphorylated Akt P Akt, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phosphorylated p akt
    Recombinant adenovirus enhancing apoptosis through PI3K/AKT-caspase-3 ameliorates resistance to DDP on SKOV3 cells. (A) The representative images of the Annexin V-APC staining assay. (B) Recombinant adenovirus and/or DDP promoted the apoptosis of ovarian cancer cells and the apoptosis rate in combination group was significantly higher than that in DDP group (n=4 in each group). (C) PI3K was significantly suppressed by recombinant adenovirus and/or DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (D) AKT was significantly suppressed by recombinant adenovirus and (or) DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (E) Procaspase-3 was significantly reduced after recombinant adenovirus and/or DDP incubation, whereas cleaved caspase-3 was considerably increased (n=4 in each group). Data are shown as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 vs. corresponding controls; # P<0.05, ## P<0.01, ### P<0.001 vs. DDP groups, & P<0.05, &&& P<0.001 vs. DDP combined with Ad-NC groups. DDP, cisplatin; si, short interfering; NC, negative control; p-. <t>phosphorylated.</t>
    Phosphorylated P Akt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated p akt/product/Cell Signaling Technology Inc
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    Santa Cruz Biotechnology phosphorylated akt p akt
    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of <t>phosphorylated-PI3K</t> (p-PI3K), PI3K, <t>phosphorylated-Akt</t> <t>(p-Akt),</t> and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Phosphorylated Akt P Akt, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated akt p akt/product/Santa Cruz Biotechnology
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    Cell Signaling Technology Inc phosphorylated akt p akt
    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of <t>phosphorylated-PI3K</t> (p-PI3K), PI3K, <t>phosphorylated-Akt</t> <t>(p-Akt),</t> and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Phosphorylated Akt P Akt, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphorylated akt p akt/product/Cell Signaling Technology Inc
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    Affinity Biosciences anti phosphorylated akt p akt
    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of <t>phosphorylated-PI3K</t> (p-PI3K), PI3K, <t>phosphorylated-Akt</t> <t>(p-Akt),</t> and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Anti Phosphorylated Akt P Akt, supplied by Affinity Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phosphorylated akt p akt/product/Affinity Biosciences
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    Danaher Inc anti phosphorylated akt p akt
    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of <t>phosphorylated-PI3K</t> (p-PI3K), PI3K, <t>phosphorylated-Akt</t> <t>(p-Akt),</t> and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).
    Anti Phosphorylated Akt P Akt, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phosphorylated akt p akt/product/Danaher Inc
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    Image Search Results


    Expression of phosphorylated-AKT (p-AKT), AKT, phosphorylated-GSK3β (p-GSK3β), GSK3β, and β-catenin in the nucleus of the ipsilateral hemisphere (excluding the infarct area). (A) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. Protein expression levels were normalized to GAPDH (total protein) or H3 (nuclear protein) ( n = 3). Expression of nuclear β-catenin, p-AKT, and p-GSK3β were higher in rats of the TMS (+) group compared with the TMS (–) group at 3, 7, 14, and 28 days. (B) Immunofluorescence of nuclear β-catenin (red, Alexa Fluor® 555) and Nestin (green)-positive cells in the TMS (+), TMS (–), and Sham groups ( n = 5). Nuclear β-catenin was upregulated in the TMS (+) group (analysis of variance [ANOVA]). Scale bars: 25 μm. (C) Ki67 (red, Alexa Fluor® 555) and Nestin (green, Alexa Fluor® 488)-positive cells in the TMS (–), TMS (H), and TMS + LY groups ( n = 5). More Ki67 + /Nestin + cells were detected in the TMS (H) group. Scale bars: 25 μm. (D) CCK8 analysis. Cell viability was decreased in the TMS (H) + LY group ( n = 5). (E) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (A: Student's t -test; B, D, E: Brown-Forsythe and Welch ANOVA tests followed by Dunnett's T3 multiple comparison test). D3/7/14/28 TMS (–): rats with no TMS at 3/7/14/28 days group; D3/7/14/28 TMS (+): rats with 10 Hz TMS at 3/7/14/28 days group; GAPDH: glyceric acid phosphated hydrogenase; ns: no significant; NSC: neural stem cell; SHAM: sham-operation group; TMS: transcranial magnetic stimulation; TMS (–): no TMS group; TMS (+): high-frequency TMS with 10 Hz group; TMS(H): NSCs with 10 Hz high-frequency TMS group; TMS(H) + LY: NSCs with 10 Hz high-frequency TMS and LY294002 group.

    Journal: Neural Regeneration Research

    Article Title: High-frequency repetitive transcranial magnetic stimulation promotes neural stem cell proliferation after ischemic stroke

    doi: 10.4103/1673-5374.389303

    Figure Lengend Snippet: Expression of phosphorylated-AKT (p-AKT), AKT, phosphorylated-GSK3β (p-GSK3β), GSK3β, and β-catenin in the nucleus of the ipsilateral hemisphere (excluding the infarct area). (A) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. Protein expression levels were normalized to GAPDH (total protein) or H3 (nuclear protein) ( n = 3). Expression of nuclear β-catenin, p-AKT, and p-GSK3β were higher in rats of the TMS (+) group compared with the TMS (–) group at 3, 7, 14, and 28 days. (B) Immunofluorescence of nuclear β-catenin (red, Alexa Fluor® 555) and Nestin (green)-positive cells in the TMS (+), TMS (–), and Sham groups ( n = 5). Nuclear β-catenin was upregulated in the TMS (+) group (analysis of variance [ANOVA]). Scale bars: 25 μm. (C) Ki67 (red, Alexa Fluor® 555) and Nestin (green, Alexa Fluor® 488)-positive cells in the TMS (–), TMS (H), and TMS + LY groups ( n = 5). More Ki67 + /Nestin + cells were detected in the TMS (H) group. Scale bars: 25 μm. (D) CCK8 analysis. Cell viability was decreased in the TMS (H) + LY group ( n = 5). (E) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (A: Student's t -test; B, D, E: Brown-Forsythe and Welch ANOVA tests followed by Dunnett's T3 multiple comparison test). D3/7/14/28 TMS (–): rats with no TMS at 3/7/14/28 days group; D3/7/14/28 TMS (+): rats with 10 Hz TMS at 3/7/14/28 days group; GAPDH: glyceric acid phosphated hydrogenase; ns: no significant; NSC: neural stem cell; SHAM: sham-operation group; TMS: transcranial magnetic stimulation; TMS (–): no TMS group; TMS (+): high-frequency TMS with 10 Hz group; TMS(H): NSCs with 10 Hz high-frequency TMS group; TMS(H) + LY: NSCs with 10 Hz high-frequency TMS and LY294002 group.

    Article Snippet: They were then incubated overnight at 4°C with primary antibodies against: rabbit anti-β-catenin (Abcam, Cat# ab32572, RRID: AB_2629234), rabbit anti-H3 (Abcam, Cat# ab1791, RRID: AB_11184913), rabbit anti-phosphorylated-GSK3β (p-GSK3β) (phospho Ser9, Abcam, Cat# ab93926, RRID: AB_2928098), rabbit anti-GSK3β (Abcam, Cat# ab280376, RRID: AB_2928097), rabbit anti-AKT (Abcam, Cat# ab188099, RRID: AB_1662415), rabbit anti-phosphorylated-AKT (p-AKT) (phospho S473, Abcam, Cat# ab81283, RRID: AB_1662951), and rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Abcam, Cat# ab181602, RRID: AB_1726478); all at a dilution of 1:500.

    Techniques: Expressing, Western Blot, Immunofluorescence, Comparison

    rTMS activates AKT through the Ca 2+ /P2 calcium channel/CaM pathway. (A) Calcium imaging of NSCs by Fluo-4 AM labeling. Cells in the TMS (+) group showed stronger signal. Scale bars: 25 μm. (B, C) Levels of Ca 2+ influx by fluorescence intensity (B; n = 5) and flow cytometry (C; n = 4). (D) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Expression of nuclear β-catenin, p-GSK3β, and p-AKT were higher in NSCs of the TMS (H) group compared with other groups. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (B: Student's t -test; D: Brown-Forsythe and Welch analysis of variance tests followed by Dunnett's T3 multiple comparison test). 2-APB: 2-Aminoethyl diphenylborinate; CPZ: chlorpromazine; CTRL: NSCs with no OGD group; NSC: neural stem cell; OGD: NSCs with OGD group; p-AKT: phosphorylated-AKT; p-GSK3β: phosphorylated-glycogen synthase kinase-3β; TMS(H): NSCs with 10 Hz high-frequency TMS group; U7: inhibitor U73122.

    Journal: Neural Regeneration Research

    Article Title: High-frequency repetitive transcranial magnetic stimulation promotes neural stem cell proliferation after ischemic stroke

    doi: 10.4103/1673-5374.389303

    Figure Lengend Snippet: rTMS activates AKT through the Ca 2+ /P2 calcium channel/CaM pathway. (A) Calcium imaging of NSCs by Fluo-4 AM labeling. Cells in the TMS (+) group showed stronger signal. Scale bars: 25 μm. (B, C) Levels of Ca 2+ influx by fluorescence intensity (B; n = 5) and flow cytometry (C; n = 4). (D) Western blot analysis of p-AKT, p-GSK3β, and β-catenin in the nucleus. GAPDH (total protein) or H3 (nuclear protein) were used as the internal control. Expression of nuclear β-catenin, p-GSK3β, and p-AKT were higher in NSCs of the TMS (H) group compared with other groups. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01 (B: Student's t -test; D: Brown-Forsythe and Welch analysis of variance tests followed by Dunnett's T3 multiple comparison test). 2-APB: 2-Aminoethyl diphenylborinate; CPZ: chlorpromazine; CTRL: NSCs with no OGD group; NSC: neural stem cell; OGD: NSCs with OGD group; p-AKT: phosphorylated-AKT; p-GSK3β: phosphorylated-glycogen synthase kinase-3β; TMS(H): NSCs with 10 Hz high-frequency TMS group; U7: inhibitor U73122.

    Article Snippet: They were then incubated overnight at 4°C with primary antibodies against: rabbit anti-β-catenin (Abcam, Cat# ab32572, RRID: AB_2629234), rabbit anti-H3 (Abcam, Cat# ab1791, RRID: AB_11184913), rabbit anti-phosphorylated-GSK3β (p-GSK3β) (phospho Ser9, Abcam, Cat# ab93926, RRID: AB_2928098), rabbit anti-GSK3β (Abcam, Cat# ab280376, RRID: AB_2928097), rabbit anti-AKT (Abcam, Cat# ab188099, RRID: AB_1662415), rabbit anti-phosphorylated-AKT (p-AKT) (phospho S473, Abcam, Cat# ab81283, RRID: AB_1662951), and rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Abcam, Cat# ab181602, RRID: AB_1726478); all at a dilution of 1:500.

    Techniques: Imaging, Labeling, Fluorescence, Flow Cytometry, Western Blot, Expressing, Comparison

    Recombinant adenovirus enhancing apoptosis through PI3K/AKT-caspase-3 ameliorates resistance to DDP on SKOV3 cells. (A) The representative images of the Annexin V-APC staining assay. (B) Recombinant adenovirus and/or DDP promoted the apoptosis of ovarian cancer cells and the apoptosis rate in combination group was significantly higher than that in DDP group (n=4 in each group). (C) PI3K was significantly suppressed by recombinant adenovirus and/or DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (D) AKT was significantly suppressed by recombinant adenovirus and (or) DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (E) Procaspase-3 was significantly reduced after recombinant adenovirus and/or DDP incubation, whereas cleaved caspase-3 was considerably increased (n=4 in each group). Data are shown as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 vs. corresponding controls; # P<0.05, ## P<0.01, ### P<0.001 vs. DDP groups, & P<0.05, &&& P<0.001 vs. DDP combined with Ad-NC groups. DDP, cisplatin; si, short interfering; NC, negative control; p-. phosphorylated.

    Journal: Molecular Medicine Reports

    Article Title: Dual‑regulated oncolytic adenovirus carrying ERCC1 ‑siRNA gene possesses potent antitumor effect on ovarian cancer cells

    doi: 10.3892/mmr.2024.13245

    Figure Lengend Snippet: Recombinant adenovirus enhancing apoptosis through PI3K/AKT-caspase-3 ameliorates resistance to DDP on SKOV3 cells. (A) The representative images of the Annexin V-APC staining assay. (B) Recombinant adenovirus and/or DDP promoted the apoptosis of ovarian cancer cells and the apoptosis rate in combination group was significantly higher than that in DDP group (n=4 in each group). (C) PI3K was significantly suppressed by recombinant adenovirus and/or DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (D) AKT was significantly suppressed by recombinant adenovirus and (or) DDP and Ad-siERCC1 could greatly enhanced the effect of DDP (n=4 in each group). (E) Procaspase-3 was significantly reduced after recombinant adenovirus and/or DDP incubation, whereas cleaved caspase-3 was considerably increased (n=4 in each group). Data are shown as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 vs. corresponding controls; # P<0.05, ## P<0.01, ### P<0.001 vs. DDP groups, & P<0.05, &&& P<0.001 vs. DDP combined with Ad-NC groups. DDP, cisplatin; si, short interfering; NC, negative control; p-. phosphorylated.

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-PI3K (1:1,000; cat. no. 4257; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (Ser473; 1:1,000; cat. no. 4060; Cell Signaling Technology, Inc.), anti-AKT (1:1,000; cat. no. ab179463; Abcam), anti-caspase-3 (1:1,000; cat. no. sc-7272; Santa Cruz Biotechnology, Inc.), cleaved caspase-3 (1:1,000; cat. no. 9661S; Cell Signaling Technology, Inc.), GAPDH (1:2,000; cat. no. sc-32233; Santa Cruz Biotechnology), β-Actin (1:2,000; cat. no. sc-8432; Santa Cruz Biotechnology).

    Techniques: Recombinant, Staining, Incubation, Negative Control

    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-PI3K (p-PI3K), PI3K, phosphorylated-Akt (p-Akt), and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Journal: Preventive Nutrition and Food Science

    Article Title: Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes

    doi: 10.3746/pnf.2024.29.2.154

    Figure Lengend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-PI3K (p-PI3K), PI3K, phosphorylated-Akt (p-Akt), and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Article Snippet: Next, the membrane was incubated overnight at 4°C with the primary antibodies against α-tubulin, NF-κB (Santa Cruz Biotechnology Inc.), PI3K, phosphorylated-PI3K (p-PI3K), Akt, phosphorylated-Akt (p-Akt), 70-kDa ribosomal protein S6 kinase (p70S6K), phosphorylated-p70S6K (p-p70S6K), eukaryotic initiation factor 4E binding protein 1 (4EBP1), phosphorylated-4EBP1 (p-4EBP1), mTOR, phosphorylated-mTOR (p-mTOR), FoxO3, and phosphorylated-FoxO3 (p-FoxO3) (Cell Signaling Technology).

    Techniques: Western Blot, Expressing, Control

    Effects of Agastache rugosa extract (ARE) and tilianin on the mammalian target of rapamycin (mTOR) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-mTOR (p-mTOR), mTOR, phosphorylated-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phosphorylated-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), and 4EBP1 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Journal: Preventive Nutrition and Food Science

    Article Title: Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes

    doi: 10.3746/pnf.2024.29.2.154

    Figure Lengend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on the mammalian target of rapamycin (mTOR) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-mTOR (p-mTOR), mTOR, phosphorylated-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phosphorylated-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), and 4EBP1 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Article Snippet: Next, the membrane was incubated overnight at 4°C with the primary antibodies against α-tubulin, NF-κB (Santa Cruz Biotechnology Inc.), PI3K, phosphorylated-PI3K (p-PI3K), Akt, phosphorylated-Akt (p-Akt), 70-kDa ribosomal protein S6 kinase (p70S6K), phosphorylated-p70S6K (p-p70S6K), eukaryotic initiation factor 4E binding protein 1 (4EBP1), phosphorylated-4EBP1 (p-4EBP1), mTOR, phosphorylated-mTOR (p-mTOR), FoxO3, and phosphorylated-FoxO3 (p-FoxO3) (Cell Signaling Technology).

    Techniques: Western Blot, Expressing, Binding Assay, Control

    Effects of Agastache rugosa extract (ARE) and tilianin on protein-degradation-related pathways in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-forkhead box class O 3 (p-FoxO3) and FoxO3 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Reverse transcription polymerase chain reaction was used to determine the mRNA expression levels of muscle RING-finger protein-1 (MuRF1) and atrogin-1 in response to (C) ARE and (D) tilianin. β-Actin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. # P <0.05, ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Journal: Preventive Nutrition and Food Science

    Article Title: Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes

    doi: 10.3746/pnf.2024.29.2.154

    Figure Lengend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on protein-degradation-related pathways in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-forkhead box class O 3 (p-FoxO3) and FoxO3 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Reverse transcription polymerase chain reaction was used to determine the mRNA expression levels of muscle RING-finger protein-1 (MuRF1) and atrogin-1 in response to (C) ARE and (D) tilianin. β-Actin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. # P <0.05, ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Article Snippet: Next, the membrane was incubated overnight at 4°C with the primary antibodies against α-tubulin, NF-κB (Santa Cruz Biotechnology Inc.), PI3K, phosphorylated-PI3K (p-PI3K), Akt, phosphorylated-Akt (p-Akt), 70-kDa ribosomal protein S6 kinase (p70S6K), phosphorylated-p70S6K (p-p70S6K), eukaryotic initiation factor 4E binding protein 1 (4EBP1), phosphorylated-4EBP1 (p-4EBP1), mTOR, phosphorylated-mTOR (p-mTOR), FoxO3, and phosphorylated-FoxO3 (p-FoxO3) (Cell Signaling Technology).

    Techniques: Western Blot, Expressing, Control, Reverse Transcription, Polymerase Chain Reaction

    Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-PI3K (p-PI3K), PI3K, phosphorylated-Akt (p-Akt), and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Journal: Preventive Nutrition and Food Science

    Article Title: Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes

    doi: 10.3746/pnf.2024.29.2.154

    Figure Lengend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-PI3K (p-PI3K), PI3K, phosphorylated-Akt (p-Akt), and Akt in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Article Snippet: Next, the membrane was incubated overnight at 4°C with the primary antibodies against α-tubulin, NF-κB (Santa Cruz Biotechnology Inc.), PI3K, phosphorylated-PI3K (p-PI3K), Akt, phosphorylated-Akt (p-Akt), 70-kDa ribosomal protein S6 kinase (p70S6K), phosphorylated-p70S6K (p-p70S6K), eukaryotic initiation factor 4E binding protein 1 (4EBP1), phosphorylated-4EBP1 (p-4EBP1), mTOR, phosphorylated-mTOR (p-mTOR), FoxO3, and phosphorylated-FoxO3 (p-FoxO3) (Cell Signaling Technology).

    Techniques: Western Blot, Expressing, Control

    Effects of Agastache rugosa extract (ARE) and tilianin on the mammalian target of rapamycin (mTOR) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-mTOR (p-mTOR), mTOR, phosphorylated-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phosphorylated-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), and 4EBP1 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Journal: Preventive Nutrition and Food Science

    Article Title: Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes

    doi: 10.3746/pnf.2024.29.2.154

    Figure Lengend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on the mammalian target of rapamycin (mTOR) pathway in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-mTOR (p-mTOR), mTOR, phosphorylated-70-kDa ribosomal protein S6 kinase (p-p70S6K), p70S6K, phosphorylated-eukaryotic initiation factor 4E binding protein 1 (p-4EBP1), and 4EBP1 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Article Snippet: Next, the membrane was incubated overnight at 4°C with the primary antibodies against α-tubulin, NF-κB (Santa Cruz Biotechnology Inc.), PI3K, phosphorylated-PI3K (p-PI3K), Akt, phosphorylated-Akt (p-Akt), 70-kDa ribosomal protein S6 kinase (p70S6K), phosphorylated-p70S6K (p-p70S6K), eukaryotic initiation factor 4E binding protein 1 (4EBP1), phosphorylated-4EBP1 (p-4EBP1), mTOR, phosphorylated-mTOR (p-mTOR), FoxO3, and phosphorylated-FoxO3 (p-FoxO3) (Cell Signaling Technology).

    Techniques: Western Blot, Expressing, Binding Assay, Control

    Effects of Agastache rugosa extract (ARE) and tilianin on protein-degradation-related pathways in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-forkhead box class O 3 (p-FoxO3) and FoxO3 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Reverse transcription polymerase chain reaction was used to determine the mRNA expression levels of muscle RING-finger protein-1 (MuRF1) and atrogin-1 in response to (C) ARE and (D) tilianin. β-Actin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. # P <0.05, ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Journal: Preventive Nutrition and Food Science

    Article Title: Korean Mint ( Agastache rugosa ) Extract and Its Bioactive Compound Tilianin Alleviate Muscle Atrophy via the PI3K/Akt/FoxO3 Pathway in C2C12 Myotubes

    doi: 10.3746/pnf.2024.29.2.154

    Figure Lengend Snippet: Effects of Agastache rugosa extract (ARE) and tilianin on protein-degradation-related pathways in C2C12 myotubes. C2C12 myotubes were simultaneously exposed to tumor necrosis factor-α (TNF-α, 50 ng/mL) along with ARE (40 and 80 μg/mL) or tilianin (20 and 40 μM). Western blot analysis was used to assess the protein expression levels of phosphorylated-forkhead box class O 3 (p-FoxO3) and FoxO3 in response to (A) ARE and (B) tilianin. α-Tubulin served as the internal control. Reverse transcription polymerase chain reaction was used to determine the mRNA expression levels of muscle RING-finger protein-1 (MuRF1) and atrogin-1 in response to (C) ARE and (D) tilianin. β-Actin served as the internal control. Experimental results were presented as the mean±SD. Significant differences were assessed using Duncan’s multiple range test. # P <0.05, ## P <0.01 (control group vs. TNF-α group); * P <0.05, ** P <0.01 (TNF-α group vs. ARE or tilianin group).

    Article Snippet: Next, the membrane was incubated overnight at 4°C with the primary antibodies against α-tubulin, NF-κB (Santa Cruz Biotechnology Inc.), PI3K, phosphorylated-PI3K (p-PI3K), Akt, phosphorylated-Akt (p-Akt), 70-kDa ribosomal protein S6 kinase (p70S6K), phosphorylated-p70S6K (p-p70S6K), eukaryotic initiation factor 4E binding protein 1 (4EBP1), phosphorylated-4EBP1 (p-4EBP1), mTOR, phosphorylated-mTOR (p-mTOR), FoxO3, and phosphorylated-FoxO3 (p-FoxO3) (Cell Signaling Technology).

    Techniques: Western Blot, Expressing, Control, Reverse Transcription, Polymerase Chain Reaction