anti akt  (Abcam)

 
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    Name:
    Anti AKT1 phospho S473 antibody EP2109Y
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    Catalog Number:
    AB81283
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    Structured Review

    Abcam anti akt
    <t>ARID1A</t> siRNA increases the expression levels of <t>AKT.</t> Western blotting was performed to quantify the expression levels of AKT. β-actin was used as a control for the expression levels. Data are presented as the mean + standard deviation (n=3). *P

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    anti akt - by Bioz Stars, 2021-06
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    Images

    1) Product Images from "ARID1A gene silencing reduces the sensitivity of ovarian clear cell carcinoma to cisplatin"

    Article Title: ARID1A gene silencing reduces the sensitivity of ovarian clear cell carcinoma to cisplatin

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2016.3863

    ARID1A siRNA increases the expression levels of AKT. Western blotting was performed to quantify the expression levels of AKT. β-actin was used as a control for the expression levels. Data are presented as the mean + standard deviation (n=3). *P
    Figure Legend Snippet: ARID1A siRNA increases the expression levels of AKT. Western blotting was performed to quantify the expression levels of AKT. β-actin was used as a control for the expression levels. Data are presented as the mean + standard deviation (n=3). *P

    Techniques Used: Expressing, Western Blot, Standard Deviation

    2) Product Images from "Resibufogenin inhibits ovarian clear cell carcinoma (OCCC) growth in vivo, and migration of OCCC cells in vitro, by down-regulating the PI3K/AKT and actin cytoskeleton signaling pathways"

    Article Title: Resibufogenin inhibits ovarian clear cell carcinoma (OCCC) growth in vivo, and migration of OCCC cells in vitro, by down-regulating the PI3K/AKT and actin cytoskeleton signaling pathways

    Journal: American Journal of Translational Research

    doi:

    KEGG pathway enrichment analyses of genes that were found to be differentially expressed upon resibufogenin treatment. A. Distribution of TOV-21G transcriptome sequences among KEGG pathways. All 13 apoptosis and migration-related pathways are shown. LgP is the negative logarithm of P -value; larger LgP values indicate smaller P -values. B. The differentially expressed genes that participate in the PI3K-AKT signaling pathway mapped in KEGG. C. The differentially expressed genes that participate in actin cytoskeleton pathway mapped in KEGG. The tree displays the log-transformed value of average fold changes. Red indicates up-regulated gene expression levels, whereas blue indicates down-regulated expression levels of genes. D. The differentially expressed genes that participated in the actin cytoskeleton signaling pathway mapped in KEGG. E. The brief graph of the differentially expressed genes that participated in the actin cytoskeleton signaling pathway mapped in KEGG.
    Figure Legend Snippet: KEGG pathway enrichment analyses of genes that were found to be differentially expressed upon resibufogenin treatment. A. Distribution of TOV-21G transcriptome sequences among KEGG pathways. All 13 apoptosis and migration-related pathways are shown. LgP is the negative logarithm of P -value; larger LgP values indicate smaller P -values. B. The differentially expressed genes that participate in the PI3K-AKT signaling pathway mapped in KEGG. C. The differentially expressed genes that participate in actin cytoskeleton pathway mapped in KEGG. The tree displays the log-transformed value of average fold changes. Red indicates up-regulated gene expression levels, whereas blue indicates down-regulated expression levels of genes. D. The differentially expressed genes that participated in the actin cytoskeleton signaling pathway mapped in KEGG. E. The brief graph of the differentially expressed genes that participated in the actin cytoskeleton signaling pathway mapped in KEGG.

    Techniques Used: Migration, Transformation Assay, Expressing

    Resibufogenin down-regulates expression of genes in the PI3K/AKT and actin cytoskeleton pathways. ES-2 and TOV-21G cells were divided into 2 groups: the control group, and the resibufogenin-treated group (20 μM). qRT-PCR was performed to detect the mRNAs levels of PI3K (A and E), MDM2 (B and F), GRLF1 (C and G), Myosin-II (D and H), RTK (I and J), and SOS (K and L) after 12 hours of treatment. *P
    Figure Legend Snippet: Resibufogenin down-regulates expression of genes in the PI3K/AKT and actin cytoskeleton pathways. ES-2 and TOV-21G cells were divided into 2 groups: the control group, and the resibufogenin-treated group (20 μM). qRT-PCR was performed to detect the mRNAs levels of PI3K (A and E), MDM2 (B and F), GRLF1 (C and G), Myosin-II (D and H), RTK (I and J), and SOS (K and L) after 12 hours of treatment. *P

    Techniques Used: Expressing, Quantitative RT-PCR

    Resibufogenin inhibits proliferation and invasion of OCCC cells in vivo through the PI3K/AKT and actin cytoskeleton pathways. Representative photomicrographs showing immunohistochemically stained xenograft tumor sections (100 × and 400 × magnification) showing PI3K, AKT and MDM2 expression.
    Figure Legend Snippet: Resibufogenin inhibits proliferation and invasion of OCCC cells in vivo through the PI3K/AKT and actin cytoskeleton pathways. Representative photomicrographs showing immunohistochemically stained xenograft tumor sections (100 × and 400 × magnification) showing PI3K, AKT and MDM2 expression.

    Techniques Used: In Vivo, Staining, Expressing

    Resibufogenin down-regulates expression of proteins in the PI3K/AKT and actin cytoskeleton pathways in vitro . ES-2 and TOV-21G cells were divided into 2 groups: the control group, and the resibufogenin-treated group (20 μM). Western blotting assay was performed to detect the protein levels of PI3K (A and B), AKT (A), MDM2 (D), and Myosin-II (D) after 24 hours of treatment. The relative expression levels of the indicated proteins (relative to the expression level of actin) were quantitated using densitometry and are depicted bar graphically in (B, C, E, and F). *P
    Figure Legend Snippet: Resibufogenin down-regulates expression of proteins in the PI3K/AKT and actin cytoskeleton pathways in vitro . ES-2 and TOV-21G cells were divided into 2 groups: the control group, and the resibufogenin-treated group (20 μM). Western blotting assay was performed to detect the protein levels of PI3K (A and B), AKT (A), MDM2 (D), and Myosin-II (D) after 24 hours of treatment. The relative expression levels of the indicated proteins (relative to the expression level of actin) were quantitated using densitometry and are depicted bar graphically in (B, C, E, and F). *P

    Techniques Used: Expressing, In Vitro, Western Blot

    Schematic diagram of the proposed mechanism by which resibufogenin inhibits proliferation and invasion in OCCC cells. Resibufogenin decreases the levels of PI3K, AKT, MDM2, and Myosin in the PI3K/AKT and actin cytoskeleton signaling pathways.
    Figure Legend Snippet: Schematic diagram of the proposed mechanism by which resibufogenin inhibits proliferation and invasion in OCCC cells. Resibufogenin decreases the levels of PI3K, AKT, MDM2, and Myosin in the PI3K/AKT and actin cytoskeleton signaling pathways.

    Techniques Used:

    3) Product Images from "Down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion injury via the miR‐335/ROCK1/AKT/GSK‐3β axis. Down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion injury via the miR‐335/ROCK1/AKT/GSK‐3β axis"

    Article Title: Down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion injury via the miR‐335/ROCK1/AKT/GSK‐3β axis. Down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion injury via the miR‐335/ROCK1/AKT/GSK‐3β axis

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.14724

    GAS5/miR‐335 manipulation regulated PTEN/PI3K‐AKT/GSK‐3β pathway and mitochondrial permeability transition pore (mPTP) opening. Scramble and si‐GAS5 transfection alone or together with miR‐335 inhibitor or inhibitor NC was transfected into cardiomyocytes. 24 h after transfection, cells were subjected to 6 h of hypoxia, followed by 3 h of reoxygenation. The phosphorylation levels of PTEN, AKT and GSK‐3β in H9c2 (A) and AC16 (B) cells were determined by Western blotting. The phosphorylation levels were expressed as the ratio of phospho‐protein expression to its corresponding total protein expression. mPTP opening was induced by CaCl 2 in H9c2 (C) and AC16 (D) cells. The decrease of optical density (OD) value reflected the extent of mPTP opening. min OD, OD value recorded at the onset of experiment (0 min); max OD, OD value recorded at the end of experiment (10 min). min/max OD is negatively associated with the extent of mPTP opening. CSA, cardiomyocytes treated with 0.2 mmol/L cyclosporin a (CSA), a mPTP opening inhibitor, were used as a positive control. Data are expressed as the mean ± standard deviation (SD) n = 3, ** P
    Figure Legend Snippet: GAS5/miR‐335 manipulation regulated PTEN/PI3K‐AKT/GSK‐3β pathway and mitochondrial permeability transition pore (mPTP) opening. Scramble and si‐GAS5 transfection alone or together with miR‐335 inhibitor or inhibitor NC was transfected into cardiomyocytes. 24 h after transfection, cells were subjected to 6 h of hypoxia, followed by 3 h of reoxygenation. The phosphorylation levels of PTEN, AKT and GSK‐3β in H9c2 (A) and AC16 (B) cells were determined by Western blotting. The phosphorylation levels were expressed as the ratio of phospho‐protein expression to its corresponding total protein expression. mPTP opening was induced by CaCl 2 in H9c2 (C) and AC16 (D) cells. The decrease of optical density (OD) value reflected the extent of mPTP opening. min OD, OD value recorded at the onset of experiment (0 min); max OD, OD value recorded at the end of experiment (10 min). min/max OD is negatively associated with the extent of mPTP opening. CSA, cardiomyocytes treated with 0.2 mmol/L cyclosporin a (CSA), a mPTP opening inhibitor, were used as a positive control. Data are expressed as the mean ± standard deviation (SD) n = 3, ** P

    Techniques Used: Permeability, Transfection, Western Blot, Expressing, Positive Control, Standard Deviation

    4) Product Images from "Palmitic Acid Promotes Virus Replication in Fish Cell by Modulating Autophagy Flux and TBK1-IRF3/7 Pathway"

    Article Title: Palmitic Acid Promotes Virus Replication in Fish Cell by Modulating Autophagy Flux and TBK1-IRF3/7 Pathway

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.01764

    Palmitic acid impacted autophagic flux in GS cells. (A) Accumulation of GFP-LC3 (green) were observed after palmitic acid treatment. GS cells were transfected with C1-EGFP-LC3 plasmid, then incubated with or without 0.4 mM palmitic acid. Cells were fixed with 4% polyformaldehyde and staining with Hoechst33342. Fluorescence was observed under the fluorescence microscope (Zeiss). (B) Representative image of phospho-Akt (Ser473) [p-Akt (ser473)], and phospho-mTOR (p-mTOR) detection was performed to verify Akt and mTOR inhibition by 1% BSA or palmitic acid (0.4 mM) treatment. β-tubulin was used as the internal control. (C) The expression levels of LC3, and p62 in cell lysates were evaluated after the incubation of 1% BSA or palmitic acid (0.4 mM) for 24 h. Western blot assay was carried out, and β-tubulin was used as the internal control. (D) GS cells were incubated with 1% BSA, 0.2 mM palmitic acid, or 0.4 mM palmitic acid for 24 h, and CQ was added for the last 4 h treatment. Western blot assay was carried out to detect the LC3-II and β-tubulin levels in cell lysate. Band intensity was calculated using Image J software, and the LC3-II protein level was presented by the ratio of LC3-II/β-tubulin. (E) Autophagic flux was measured. Briefly, after measuring the LC3-II protein level (LC3-II/β-tubulin) in each group, the histogram was made referring to the ratio of LC3-II level in cells treated with CQ to that of untreated cells. Setting the ratio in 1% BSA treated group as 1-fold. The data are represented as mean ± SD, and the statistical significances were determined with Student's t -test, n = 3. The significance level was defined as * p
    Figure Legend Snippet: Palmitic acid impacted autophagic flux in GS cells. (A) Accumulation of GFP-LC3 (green) were observed after palmitic acid treatment. GS cells were transfected with C1-EGFP-LC3 plasmid, then incubated with or without 0.4 mM palmitic acid. Cells were fixed with 4% polyformaldehyde and staining with Hoechst33342. Fluorescence was observed under the fluorescence microscope (Zeiss). (B) Representative image of phospho-Akt (Ser473) [p-Akt (ser473)], and phospho-mTOR (p-mTOR) detection was performed to verify Akt and mTOR inhibition by 1% BSA or palmitic acid (0.4 mM) treatment. β-tubulin was used as the internal control. (C) The expression levels of LC3, and p62 in cell lysates were evaluated after the incubation of 1% BSA or palmitic acid (0.4 mM) for 24 h. Western blot assay was carried out, and β-tubulin was used as the internal control. (D) GS cells were incubated with 1% BSA, 0.2 mM palmitic acid, or 0.4 mM palmitic acid for 24 h, and CQ was added for the last 4 h treatment. Western blot assay was carried out to detect the LC3-II and β-tubulin levels in cell lysate. Band intensity was calculated using Image J software, and the LC3-II protein level was presented by the ratio of LC3-II/β-tubulin. (E) Autophagic flux was measured. Briefly, after measuring the LC3-II protein level (LC3-II/β-tubulin) in each group, the histogram was made referring to the ratio of LC3-II level in cells treated with CQ to that of untreated cells. Setting the ratio in 1% BSA treated group as 1-fold. The data are represented as mean ± SD, and the statistical significances were determined with Student's t -test, n = 3. The significance level was defined as * p

    Techniques Used: Transfection, Plasmid Preparation, Incubation, Staining, Fluorescence, Microscopy, Inhibition, Expressing, Western Blot, Software

    5) Product Images from "Long Non-Coding RNA (lncRNA) Urothelial Carcinoma-Associated 1 (UCA1) Enhances Tamoxifen Resistance in Breast Cancer Cells via Inhibiting mTOR Signaling Pathway"

    Article Title: Long Non-Coding RNA (lncRNA) Urothelial Carcinoma-Associated 1 (UCA1) Enhances Tamoxifen Resistance in Breast Cancer Cells via Inhibiting mTOR Signaling Pathway

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

    doi: 10.12659/MSM.900689

    UCA1 activates mTOR signaling pathway in breast cancer cells. ( A–D ) Western blot analysis images ( A, B ) and quantitation of the relative gray scale ( C, D ) of the expression of p-AKT and p-mTOR in MCF-7, LCC2 and LCC9 cells ( A, C ) and in LCC2 and LCC9 cells with or without transfection of UCA1 siRNA ( B, D ). ** p
    Figure Legend Snippet: UCA1 activates mTOR signaling pathway in breast cancer cells. ( A–D ) Western blot analysis images ( A, B ) and quantitation of the relative gray scale ( C, D ) of the expression of p-AKT and p-mTOR in MCF-7, LCC2 and LCC9 cells ( A, C ) and in LCC2 and LCC9 cells with or without transfection of UCA1 siRNA ( B, D ). ** p

    Techniques Used: Western Blot, Quantitation Assay, Expressing, Transfection

    6) Product Images from "MiR-4766-5p Inhibits The Development And Progression Of Gastric Cancer By Targeting NKAP"

    Article Title: MiR-4766-5p Inhibits The Development And Progression Of Gastric Cancer By Targeting NKAP

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S220234

    MiR-4766-5p inhibited the activation of AKT/mTOR pathways. ( A ) Western blots for the protein expression in different cells lines for AGS and MKN45, respectively. ( B ) Fold change for AGS in different cell lines. ( C ) Fold change for MKN45 in different cell lines. *P
    Figure Legend Snippet: MiR-4766-5p inhibited the activation of AKT/mTOR pathways. ( A ) Western blots for the protein expression in different cells lines for AGS and MKN45, respectively. ( B ) Fold change for AGS in different cell lines. ( C ) Fold change for MKN45 in different cell lines. *P

    Techniques Used: Activation Assay, Western Blot, Expressing

    7) Product Images from "MiR-195-5p Ameliorates Cerebral Ischemia-Reperfusion Injury by Regulating the PTEN-AKT Signaling Pathway"

    Article Title: MiR-195-5p Ameliorates Cerebral Ischemia-Reperfusion Injury by Regulating the PTEN-AKT Signaling Pathway

    Journal: Neuropsychiatric Disease and Treatment

    doi: 10.2147/NDT.S297975

    Overexpression of miR-195-5p ameliorated cerebral I/R injury in vivo. MiR-195-5p agomir or agomir NC (100 μM) was injected into mice, and mice then received 2 h of MCAO followed by 24 h of reperfusion. ( A ) The expression of miR-195-5p in brain tissues was evaluated by qRT-PCR. ( B ) The infarct volume in brains was assessed by TCC staining. ( C ) H E staining of whole brain in mice. Scare bar = 400 μm. ( D ) The apoptosis of cortical neurons was evaluated by TUNEL staining. TUNEL positive cells (green) and DAPI positive cells (blue), and images were merged by using fluorescence microscope. Scare bar = 50 μm. ( E ) Neurological deficit score. ( F ) The expression of PTEN, p-AKT and AKT in brain tissues was evaluated by Western blot. Data were presented as means ± SD. ** p
    Figure Legend Snippet: Overexpression of miR-195-5p ameliorated cerebral I/R injury in vivo. MiR-195-5p agomir or agomir NC (100 μM) was injected into mice, and mice then received 2 h of MCAO followed by 24 h of reperfusion. ( A ) The expression of miR-195-5p in brain tissues was evaluated by qRT-PCR. ( B ) The infarct volume in brains was assessed by TCC staining. ( C ) H E staining of whole brain in mice. Scare bar = 400 μm. ( D ) The apoptosis of cortical neurons was evaluated by TUNEL staining. TUNEL positive cells (green) and DAPI positive cells (blue), and images were merged by using fluorescence microscope. Scare bar = 50 μm. ( E ) Neurological deficit score. ( F ) The expression of PTEN, p-AKT and AKT in brain tissues was evaluated by Western blot. Data were presented as means ± SD. ** p

    Techniques Used: Over Expression, In Vivo, Injection, Mouse Assay, Expressing, Quantitative RT-PCR, Staining, TUNEL Assay, Fluorescence, Microscopy, Western Blot

    Overexpression of miR-195-5p improved I/R injury in vitro by regulating the PTEN-AKT signaling pathway. HBMVECs were transfected with miR-195-5p mimics, or co-transfected with miR-195-5p mimics and pc-PTEN, and then induced by 2 h of OGD followed by 24 h of re-oxygenation. ( A ) Cell viability was evaluated by CCK-8 assay. ( B ) The release of LDH was detected by specific cytotoxicity assay kit. ( C ) The expression of PTEN, AKT and p-AKT was detected by Western blot. ( D ) Cell apoptosis was evaluated by flow cytometer. ( E ) The protein expression of apoptosis-related makers was detected by Western blot. Data were presented as means ± SD. * p
    Figure Legend Snippet: Overexpression of miR-195-5p improved I/R injury in vitro by regulating the PTEN-AKT signaling pathway. HBMVECs were transfected with miR-195-5p mimics, or co-transfected with miR-195-5p mimics and pc-PTEN, and then induced by 2 h of OGD followed by 24 h of re-oxygenation. ( A ) Cell viability was evaluated by CCK-8 assay. ( B ) The release of LDH was detected by specific cytotoxicity assay kit. ( C ) The expression of PTEN, AKT and p-AKT was detected by Western blot. ( D ) Cell apoptosis was evaluated by flow cytometer. ( E ) The protein expression of apoptosis-related makers was detected by Western blot. Data were presented as means ± SD. * p

    Techniques Used: Over Expression, In Vitro, Transfection, CCK-8 Assay, Cytotoxicity Assay, Expressing, Western Blot, Flow Cytometry

    8) Product Images from "FGFR3b Extracellular Loop Mutation Lacks Tumorigenicity In Vivo but Collaborates with p53/pRB Deficiency to Induce High-grade Papillary Urothelial Carcinoma"

    Article Title: FGFR3b Extracellular Loop Mutation Lacks Tumorigenicity In Vivo but Collaborates with p53/pRB Deficiency to Induce High-grade Papillary Urothelial Carcinoma

    Journal: Scientific Reports

    doi: 10.1038/srep25596

    Morphological and molecular characterization of transgenic mice expressing FGFR3b-S243C. ( A ) Representative H E-stained images of urothelial cross-sections from un-induced double transgenic mice (UI, 6 months old) and induced (I) double transgenic mice (6 and 12 months old). Note the complete absence of urothelial abnormality in the induced mice. Magnification, 200x. ( B ) Western blotting showing the upregulation of phosphorylated MAPK and AKT in induced mice, as compared with uninduced mice (3 mice each condition). ( C , D ) Real-time PCR and Western blotting showing the upregulation of p16-pRb and p19-p53-p21 tumor suppressor pathway components. N = 5 in ( C ).
    Figure Legend Snippet: Morphological and molecular characterization of transgenic mice expressing FGFR3b-S243C. ( A ) Representative H E-stained images of urothelial cross-sections from un-induced double transgenic mice (UI, 6 months old) and induced (I) double transgenic mice (6 and 12 months old). Note the complete absence of urothelial abnormality in the induced mice. Magnification, 200x. ( B ) Western blotting showing the upregulation of phosphorylated MAPK and AKT in induced mice, as compared with uninduced mice (3 mice each condition). ( C , D ) Real-time PCR and Western blotting showing the upregulation of p16-pRb and p19-p53-p21 tumor suppressor pathway components. N = 5 in ( C ).

    Techniques Used: Transgenic Assay, Mouse Assay, Expressing, Staining, Western Blot, Real-time Polymerase Chain Reaction

    Signaling pathway status in triple transgenic mice expressing both FGFR3b-S243C and SV40T. Paraffin-embedded sections from the bladders of transgenic mouse groups shown in Fig. 5 were processed routinely for immunohistochemical staining using the antibodies indicated on the left. Note the marked upregulation of both MAPK and AKT pathways, particularly phosphorylated S6 and phosphorylated MAPK in triple transgenic mice treated with doxycycline (the right most panel). Magnification, 200x.
    Figure Legend Snippet: Signaling pathway status in triple transgenic mice expressing both FGFR3b-S243C and SV40T. Paraffin-embedded sections from the bladders of transgenic mouse groups shown in Fig. 5 were processed routinely for immunohistochemical staining using the antibodies indicated on the left. Note the marked upregulation of both MAPK and AKT pathways, particularly phosphorylated S6 and phosphorylated MAPK in triple transgenic mice treated with doxycycline (the right most panel). Magnification, 200x.

    Techniques Used: Transgenic Assay, Mouse Assay, Expressing, Immunohistochemistry, Staining

    9) Product Images from "SIL1 functions as an oncogene in glioma by AKT/mTOR signaling pathway"

    Article Title: SIL1 functions as an oncogene in glioma by AKT/mTOR signaling pathway

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S167552

    Downregulation of SIL1 inhibited AKT/mTOR signaling pathway. ( A ) Western blot image and ( B ) quantification analysis indicated that siSIL1 reduced the phosphorylation level of AKT and mTOR without affecting protein expression, as well as decreasing expression of the downstream effector p70S6K. Protein expression was normalized to siNC group. Note: * P
    Figure Legend Snippet: Downregulation of SIL1 inhibited AKT/mTOR signaling pathway. ( A ) Western blot image and ( B ) quantification analysis indicated that siSIL1 reduced the phosphorylation level of AKT and mTOR without affecting protein expression, as well as decreasing expression of the downstream effector p70S6K. Protein expression was normalized to siNC group. Note: * P

    Techniques Used: Western Blot, Expressing

    10) Product Images from "Human amniotic mesenchymal stem cells inhibit hepatocellular carcinoma in tumour‐bearing mice, et al. Human amniotic mesenchymal stem cells inhibit hepatocellular carcinoma in tumour‐bearing mice"

    Article Title: Human amniotic mesenchymal stem cells inhibit hepatocellular carcinoma in tumour‐bearing mice, et al. Human amniotic mesenchymal stem cells inhibit hepatocellular carcinoma in tumour‐bearing mice

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.15668

    DKK‐3, DKK‐1 and IGFBP‐3 derived from hAMSCs inhibit survival of Hepg2 cells through blocking Wnt/β‐catenin and IGF‐1R/PI3K/AKT pathway. A, Immunoblot analysis was performed on normal medium (Control), CM, CM + DKK‐1 antibody, CM + DKK‐3 antibody and CM + IGFBP‐3 antibody‐treated Hepg2 cell lysate using antibodies against GAPDH, Bax and Bcl‐2. B, Hepg2 cells were co‐cultured with normal medium (Control), hAMSC‐siMOCK, hAMSC‐siDKK‐1 and hAMSC‐siDKK‐3. Western blot analysis showed that the expression level of Bcl‐2 was increased and Bax was decreased in hAMSC‐siDKK‐3 group and hAMSC‐siDKK‐1 group when compared with hAMSC‐siMOCK group. C, Quantitative analysis of the expressions of Bcl‐2 and Bax in Hepg2 cells of different groups as in (A). D, Quantitative analysis of the expressions of Bcl‐2 and Bax in Hepg2 cells of different groups as in (B). E, Hepg2 cells were co‐cultured with normal medium (Control), hAMSC‐siMOCK and hAMSC‐siIGFBP‐3. The expression levels of Bax, Bcl‐2, P‐AKT, AKT, P‐PI3K, PI3K, P‐IGF‐1R, IGF‐1R and GAPDH in Hepg2 cells of different groups were determined by Western blot analysis. F, Quantitative analysis of the expressions of P‐IGF‐1R, P‐PI3K, P‐AKT, Bcl‐2 and Bax in Hepg2 cells of different groups as in (E). G, Schematic diagram of the extracellular and intracellular mechanisms of DKK‐3, DKK‐1 and IGFBP3 effect on the apoptosis and proliferation of Hepg2 cells. DKK‐3 and DKK‐1 secreted by hAMSCs can inhibit Wnt/β‐catenin signalling by sequestering LRP5/6, triggering apoptosis and inhibiting cell growth; IGFBP‐3 secreted by hAMSCs can inhibit IGF1 signalling by sequestering IGF1, resulting in cell apoptosis
    Figure Legend Snippet: DKK‐3, DKK‐1 and IGFBP‐3 derived from hAMSCs inhibit survival of Hepg2 cells through blocking Wnt/β‐catenin and IGF‐1R/PI3K/AKT pathway. A, Immunoblot analysis was performed on normal medium (Control), CM, CM + DKK‐1 antibody, CM + DKK‐3 antibody and CM + IGFBP‐3 antibody‐treated Hepg2 cell lysate using antibodies against GAPDH, Bax and Bcl‐2. B, Hepg2 cells were co‐cultured with normal medium (Control), hAMSC‐siMOCK, hAMSC‐siDKK‐1 and hAMSC‐siDKK‐3. Western blot analysis showed that the expression level of Bcl‐2 was increased and Bax was decreased in hAMSC‐siDKK‐3 group and hAMSC‐siDKK‐1 group when compared with hAMSC‐siMOCK group. C, Quantitative analysis of the expressions of Bcl‐2 and Bax in Hepg2 cells of different groups as in (A). D, Quantitative analysis of the expressions of Bcl‐2 and Bax in Hepg2 cells of different groups as in (B). E, Hepg2 cells were co‐cultured with normal medium (Control), hAMSC‐siMOCK and hAMSC‐siIGFBP‐3. The expression levels of Bax, Bcl‐2, P‐AKT, AKT, P‐PI3K, PI3K, P‐IGF‐1R, IGF‐1R and GAPDH in Hepg2 cells of different groups were determined by Western blot analysis. F, Quantitative analysis of the expressions of P‐IGF‐1R, P‐PI3K, P‐AKT, Bcl‐2 and Bax in Hepg2 cells of different groups as in (E). G, Schematic diagram of the extracellular and intracellular mechanisms of DKK‐3, DKK‐1 and IGFBP3 effect on the apoptosis and proliferation of Hepg2 cells. DKK‐3 and DKK‐1 secreted by hAMSCs can inhibit Wnt/β‐catenin signalling by sequestering LRP5/6, triggering apoptosis and inhibiting cell growth; IGFBP‐3 secreted by hAMSCs can inhibit IGF1 signalling by sequestering IGF1, resulting in cell apoptosis

    Techniques Used: Derivative Assay, Blocking Assay, Cell Culture, Western Blot, Expressing

    hAMSC‐derived DKK‐3, DKK‐1 and IGFBP‐3 reduced activation of Wnt/β‐catenin and IGF‐1R/PI3K/AKT signalling pathway of Hepg2 cells. A, Representative array images are shown (n = 4). DKK‐3, DKK‐1 and IGFBP‐3 are highlighted with red boxes. B, Western blot analysis of DKK‐3, DKK‐1 and IGFBP‐3 protein levels in hAMSCs and Hepg2 cells. C, Quantitative analysis of the expression of IGFBP‐3, DKK‐1 and DKK‐3 in hAMSCs and Hepg2 cells as in (B). D, Hepg2 cells were treated with normal medium (control), hAMSCs and hAMSC‐CM. Western blot analysis of protein levels of β‐catenin, GSK3β and P‐GSK3β in Hepg2 cells of each treatment group. E, Western blot analysis of protein levels of IGF‐1R, P‐ IGF‐1R, PI3K, P‐ PI3K, AKT and P‐AKT in Hepg2 cells of control, hAMSCs and hAMSC‐CM group. F, Quantitative analysis of the expression of β‐catenin and P‐GSK3β in Hepg2 cells of different groups as in (D). G, Quantitative analysis of the expression of P‐ IGF‐1R, P‐PI3K and P‐AKT in Hepg2 cells of different groups as in (E)
    Figure Legend Snippet: hAMSC‐derived DKK‐3, DKK‐1 and IGFBP‐3 reduced activation of Wnt/β‐catenin and IGF‐1R/PI3K/AKT signalling pathway of Hepg2 cells. A, Representative array images are shown (n = 4). DKK‐3, DKK‐1 and IGFBP‐3 are highlighted with red boxes. B, Western blot analysis of DKK‐3, DKK‐1 and IGFBP‐3 protein levels in hAMSCs and Hepg2 cells. C, Quantitative analysis of the expression of IGFBP‐3, DKK‐1 and DKK‐3 in hAMSCs and Hepg2 cells as in (B). D, Hepg2 cells were treated with normal medium (control), hAMSCs and hAMSC‐CM. Western blot analysis of protein levels of β‐catenin, GSK3β and P‐GSK3β in Hepg2 cells of each treatment group. E, Western blot analysis of protein levels of IGF‐1R, P‐ IGF‐1R, PI3K, P‐ PI3K, AKT and P‐AKT in Hepg2 cells of control, hAMSCs and hAMSC‐CM group. F, Quantitative analysis of the expression of β‐catenin and P‐GSK3β in Hepg2 cells of different groups as in (D). G, Quantitative analysis of the expression of P‐ IGF‐1R, P‐PI3K and P‐AKT in Hepg2 cells of different groups as in (E)

    Techniques Used: Derivative Assay, Activation Assay, Western Blot, Expressing

    11) Product Images from "SH2B1 promotes NSCLC cell proliferation through PI3K/Akt/mTOR signaling cascade"

    Article Title: SH2B1 promotes NSCLC cell proliferation through PI3K/Akt/mTOR signaling cascade

    Journal: Cancer Cell International

    doi: 10.1186/s12935-018-0632-x

    SH2B1 regulates NSCLC cell proliferation through Akt/mTOR pathway in vitro. a , b The effect of SH2B1 on Akt/mTOR signaling. a The protein levels of SH2B1, p-Akt, p-mTOR and PTEN in different panels of A549 and H1299 cells were assayed by Western blotting. b The protein levels of Akt/mTOR pathway targets (p-Akt, p-mTOR and PTEN) in different panels of H1299, which were subjected or not subjected to a Akt inhibitor, 10 μm/l deguelin for 24H, were assayed by Western blotting. Experiments were repeated three times and representative pictures are shown for a and b . GAPDH is used as an internal control. c CCK8 assay indicated that deguelin impaired the effect of SH2B1 on cell proliferation. Results are presented as mean ± SD
    Figure Legend Snippet: SH2B1 regulates NSCLC cell proliferation through Akt/mTOR pathway in vitro. a , b The effect of SH2B1 on Akt/mTOR signaling. a The protein levels of SH2B1, p-Akt, p-mTOR and PTEN in different panels of A549 and H1299 cells were assayed by Western blotting. b The protein levels of Akt/mTOR pathway targets (p-Akt, p-mTOR and PTEN) in different panels of H1299, which were subjected or not subjected to a Akt inhibitor, 10 μm/l deguelin for 24H, were assayed by Western blotting. Experiments were repeated three times and representative pictures are shown for a and b . GAPDH is used as an internal control. c CCK8 assay indicated that deguelin impaired the effect of SH2B1 on cell proliferation. Results are presented as mean ± SD

    Techniques Used: In Vitro, Western Blot, CCK-8 Assay

    Clinical relevance of SH2B1 and its targets in NSCLC. a Representative expression levels of SH2B1, p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN in NSCLC by IHC staining. Scale bars, 10 μm. b Percentages of specimens showing high or low SH2B1 expression relative to levels of p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN (all P
    Figure Legend Snippet: Clinical relevance of SH2B1 and its targets in NSCLC. a Representative expression levels of SH2B1, p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN in NSCLC by IHC staining. Scale bars, 10 μm. b Percentages of specimens showing high or low SH2B1 expression relative to levels of p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN (all P

    Techniques Used: Expressing, Immunohistochemistry, Staining

    12) Product Images from "Thyrotropin Regulates eNOS Expression in the Endothelium by PGRN Through Akt Pathway"

    Article Title: Thyrotropin Regulates eNOS Expression in the Endothelium by PGRN Through Akt Pathway

    Journal: Frontiers in Endocrinology

    doi: 10.3389/fendo.2018.00353

    Role of Akt in the induction by TSH of eNOS in HUVECs. (A) Protein expression of Akt in HUVECs stimulated by different concentrations of TSH (0, 0.1, 1, 10, and 100 mIU/ml) for 24 h; (B) Inhibition of Akt: HUVECs were pretreated with 1 μM of MK-2206 for 6 h or not, and then together with or without TSH (10 mIU/ml) for 24 h. Data were obtained from three separate experiments. HUVECs without stimulating with TSH (0 mIU/ml) served as control; M represented MK-2206; * P
    Figure Legend Snippet: Role of Akt in the induction by TSH of eNOS in HUVECs. (A) Protein expression of Akt in HUVECs stimulated by different concentrations of TSH (0, 0.1, 1, 10, and 100 mIU/ml) for 24 h; (B) Inhibition of Akt: HUVECs were pretreated with 1 μM of MK-2206 for 6 h or not, and then together with or without TSH (10 mIU/ml) for 24 h. Data were obtained from three separate experiments. HUVECs without stimulating with TSH (0 mIU/ml) served as control; M represented MK-2206; * P

    Techniques Used: Expressing, Inhibition

    Proposed mechanism of endothelial dysfunction induced by TSH. TSH up-regulated eNOS expression in HUVECs by PGRN through Akt pathway. Reduction in NO production and increase in superoxide anion indicated uncoupled eNOS.
    Figure Legend Snippet: Proposed mechanism of endothelial dysfunction induced by TSH. TSH up-regulated eNOS expression in HUVECs by PGRN through Akt pathway. Reduction in NO production and increase in superoxide anion indicated uncoupled eNOS.

    Techniques Used: Expressing

    13) Product Images from "MiR-338-3p inhibits epithelial-mesenchymal transition in gastric cancer cells by targeting ZEB2 and MACC1/Met/Akt signaling"

    Article Title: MiR-338-3p inhibits epithelial-mesenchymal transition in gastric cancer cells by targeting ZEB2 and MACC1/Met/Akt signaling

    Journal: Oncotarget

    doi:

    MiR-338-3p suppresses EMT via MACC1/Met/Akt signaling ( A ) MACC1 is predicted as the downstream target of miR-338-3p. The luciferase reporter of the miR-338-3p binding site on the wild-type and mutant MACC1 3′UTR. The replaced site is underlined. ( B ) Relative luciferase activity analyses. The vector, psiCHECK2-wMACC1, or psiCHECK2-mMACC1 plasmid (20 nM or 50 nM) was transfected into AGS and MKN-28 cells with or without miR-338-3p mimic. ( C ) MACC1, Met, pGSK-3β, GSK-3β, pAkt and Akt protein expression levels in AGS and MKN-28 cells transfected with miR-338-3p mimic, inhibitor, or the corresponding control plasmids. ( D ) Restoring MACC1 expression increased the protein expression of MACC1, Met, pGSK-3β, and pAkt in the miR-338-3p mimic-transfected cells ( E ) MiR-338-3p decreased MACC1 and vimentin expression and increased E-cadherin expression, whereas the reintroduction of MACC1 reversed these alterations. ( F ) Restoration of MACC1 re-enhanced miR-338-3p-dimished invasiveness in AGS and MKN-28 cells. ** P
    Figure Legend Snippet: MiR-338-3p suppresses EMT via MACC1/Met/Akt signaling ( A ) MACC1 is predicted as the downstream target of miR-338-3p. The luciferase reporter of the miR-338-3p binding site on the wild-type and mutant MACC1 3′UTR. The replaced site is underlined. ( B ) Relative luciferase activity analyses. The vector, psiCHECK2-wMACC1, or psiCHECK2-mMACC1 plasmid (20 nM or 50 nM) was transfected into AGS and MKN-28 cells with or without miR-338-3p mimic. ( C ) MACC1, Met, pGSK-3β, GSK-3β, pAkt and Akt protein expression levels in AGS and MKN-28 cells transfected with miR-338-3p mimic, inhibitor, or the corresponding control plasmids. ( D ) Restoring MACC1 expression increased the protein expression of MACC1, Met, pGSK-3β, and pAkt in the miR-338-3p mimic-transfected cells ( E ) MiR-338-3p decreased MACC1 and vimentin expression and increased E-cadherin expression, whereas the reintroduction of MACC1 reversed these alterations. ( F ) Restoration of MACC1 re-enhanced miR-338-3p-dimished invasiveness in AGS and MKN-28 cells. ** P

    Techniques Used: Luciferase, Binding Assay, Mutagenesis, Activity Assay, Plasmid Preparation, Transfection, Expressing

    The EMT suppressive effect induced by miR-338-3p is confirmed in GC tissues ( A , B , and C ) Western blotting assay was performed in human GC tissues (n = 7). MiR-338-3p was inversely correlated with (B) MACC1 and (C) E-cadherin protein expression in GC tissues. ( D ) ISH study of miR-338-3p and IHC analysis of ZEB2, MACC1, N-cadherin, and vimentin was performed in 20 human GC tissues. MiR-338-3p expression was inversely associated with MACC1, ZEB2, and N-cadherin. Two representative cases are shown. ( E ) Schematic representation of this study. Both ZEB2 and MACC1 are directly targeted by miR-338-3p in GC. MACC1 suppression further inhibited its downstream Met/Akt signaling and suppressed pGSK-3β phosphorylation. These effects stabilized the epithelial marker, E-cadherin and inhibited the mesenchymal marker, N-cadherin and vimentin, which resulted in the inhibition of EMT. Therefore, the findings of the present study suggests that miR-338-3p is a microRNA against GC cell migration and invasion via its EMT modulating effect.
    Figure Legend Snippet: The EMT suppressive effect induced by miR-338-3p is confirmed in GC tissues ( A , B , and C ) Western blotting assay was performed in human GC tissues (n = 7). MiR-338-3p was inversely correlated with (B) MACC1 and (C) E-cadherin protein expression in GC tissues. ( D ) ISH study of miR-338-3p and IHC analysis of ZEB2, MACC1, N-cadherin, and vimentin was performed in 20 human GC tissues. MiR-338-3p expression was inversely associated with MACC1, ZEB2, and N-cadherin. Two representative cases are shown. ( E ) Schematic representation of this study. Both ZEB2 and MACC1 are directly targeted by miR-338-3p in GC. MACC1 suppression further inhibited its downstream Met/Akt signaling and suppressed pGSK-3β phosphorylation. These effects stabilized the epithelial marker, E-cadherin and inhibited the mesenchymal marker, N-cadherin and vimentin, which resulted in the inhibition of EMT. Therefore, the findings of the present study suggests that miR-338-3p is a microRNA against GC cell migration and invasion via its EMT modulating effect.

    Techniques Used: Western Blot, Expressing, In Situ Hybridization, Immunohistochemistry, Marker, Inhibition, Migration

    14) Product Images from "Essential Role Of High Glucose-Induced Overexpression Of PKCβ And PKCδ In GLP-1 Resistance In Rodent Cardiomyocytes"

    Article Title: Essential Role Of High Glucose-Induced Overexpression Of PKCβ And PKCδ In GLP-1 Resistance In Rodent Cardiomyocytes

    Journal: Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy

    doi: 10.2147/DMSO.S215789

    Overexpression of PKCδ induced by high glucose-impaired post-receptor anti-apoptotic signaling pathways of GLP-1 in H9C2 cells by inhibition of Akt phosphorylation. ( A ) Impact of PKCα, -β, -γ, -δ siRNA and UNC siRNA on Bcl-2 and BAX mRNA level after H/R in H9C2 cells cultured by high glucose. *P
    Figure Legend Snippet: Overexpression of PKCδ induced by high glucose-impaired post-receptor anti-apoptotic signaling pathways of GLP-1 in H9C2 cells by inhibition of Akt phosphorylation. ( A ) Impact of PKCα, -β, -γ, -δ siRNA and UNC siRNA on Bcl-2 and BAX mRNA level after H/R in H9C2 cells cultured by high glucose. *P

    Techniques Used: Over Expression, Inhibition, Cell Culture

    15) Product Images from "SH2B1 promotes NSCLC cell proliferation through PI3K/Akt/mTOR signaling cascade"

    Article Title: SH2B1 promotes NSCLC cell proliferation through PI3K/Akt/mTOR signaling cascade

    Journal: Cancer Cell International

    doi: 10.1186/s12935-018-0632-x

    SH2B1 regulates NSCLC cell proliferation through Akt/mTOR pathway in vitro. a , b The effect of SH2B1 on Akt/mTOR signaling. a The protein levels of SH2B1, p-Akt, p-mTOR and PTEN in different panels of A549 and H1299 cells were assayed by Western blotting. b The protein levels of Akt/mTOR pathway targets (p-Akt, p-mTOR and PTEN) in different panels of H1299, which were subjected or not subjected to a Akt inhibitor, 10 μm/l deguelin for 24H, were assayed by Western blotting. Experiments were repeated three times and representative pictures are shown for a and b . GAPDH is used as an internal control. c CCK8 assay indicated that deguelin impaired the effect of SH2B1 on cell proliferation. Results are presented as mean ± SD
    Figure Legend Snippet: SH2B1 regulates NSCLC cell proliferation through Akt/mTOR pathway in vitro. a , b The effect of SH2B1 on Akt/mTOR signaling. a The protein levels of SH2B1, p-Akt, p-mTOR and PTEN in different panels of A549 and H1299 cells were assayed by Western blotting. b The protein levels of Akt/mTOR pathway targets (p-Akt, p-mTOR and PTEN) in different panels of H1299, which were subjected or not subjected to a Akt inhibitor, 10 μm/l deguelin for 24H, were assayed by Western blotting. Experiments were repeated three times and representative pictures are shown for a and b . GAPDH is used as an internal control. c CCK8 assay indicated that deguelin impaired the effect of SH2B1 on cell proliferation. Results are presented as mean ± SD

    Techniques Used: In Vitro, Western Blot, CCK-8 Assay

    Clinical relevance of SH2B1 and its targets in NSCLC. a Representative expression levels of SH2B1, p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN in NSCLC by IHC staining. Scale bars, 10 μm. b Percentages of specimens showing high or low SH2B1 expression relative to levels of p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN (all P
    Figure Legend Snippet: Clinical relevance of SH2B1 and its targets in NSCLC. a Representative expression levels of SH2B1, p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN in NSCLC by IHC staining. Scale bars, 10 μm. b Percentages of specimens showing high or low SH2B1 expression relative to levels of p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN (all P

    Techniques Used: Expressing, Immunohistochemistry, Staining

    16) Product Images from "Sphingosine kinase 1 is overexpressed and promotes adrenocortical carcinoma progression"

    Article Title: Sphingosine kinase 1 is overexpressed and promotes adrenocortical carcinoma progression

    Journal: Oncotarget

    doi: 10.18632/oncotarget.6564

    Effect of FTY720 on the activities of major signaling pathways, and effect of FTY720/mitotane combination on the proliferation of ACC cells ( A ) The antibodies against PI3K, phospho-PI3K Y607 (p-PI3K), AKT, phospho-AKT S473 (p-AKT), ERK, phospho-ERK pT202/pY204 + pT185/pY187 (p-ERK) were used to determine the effect of FTY720 on the activities of PI3K/Akt and MAPK signaling. Western blotting analysis revealed that FTY720 significantly reduced the levels of p-PI3K, p-AKT and p-ERK in a dose dependently manner in H295R cells; ( B ) Western blotting analysis revealed that FTY720 significantly reduced the levels of p-PI3K, p-AKT and p-ERK in a dose dependently manner in SW13 cells; ( C ) Cell viability after single or combined mitotane and FTY720 treatment. Cytotoxic response to mitotane and the combination of mitotane and FTY720 in H295R and SW13 cell lines.; ( D ) The combination of mitotane and FTY720 showed a significant synergistic anti-proliferative effect (CI = 0.90 ± 0.08) in SW13 cells that potentiated the cytotoxic effect observed by using FTY720 alone.
    Figure Legend Snippet: Effect of FTY720 on the activities of major signaling pathways, and effect of FTY720/mitotane combination on the proliferation of ACC cells ( A ) The antibodies against PI3K, phospho-PI3K Y607 (p-PI3K), AKT, phospho-AKT S473 (p-AKT), ERK, phospho-ERK pT202/pY204 + pT185/pY187 (p-ERK) were used to determine the effect of FTY720 on the activities of PI3K/Akt and MAPK signaling. Western blotting analysis revealed that FTY720 significantly reduced the levels of p-PI3K, p-AKT and p-ERK in a dose dependently manner in H295R cells; ( B ) Western blotting analysis revealed that FTY720 significantly reduced the levels of p-PI3K, p-AKT and p-ERK in a dose dependently manner in SW13 cells; ( C ) Cell viability after single or combined mitotane and FTY720 treatment. Cytotoxic response to mitotane and the combination of mitotane and FTY720 in H295R and SW13 cell lines.; ( D ) The combination of mitotane and FTY720 showed a significant synergistic anti-proliferative effect (CI = 0.90 ± 0.08) in SW13 cells that potentiated the cytotoxic effect observed by using FTY720 alone.

    Techniques Used: Western Blot

    17) Product Images from "Mast cell exosomes promote lung adenocarcinoma cell proliferation – role of KIT-stem cell factor signaling"

    Article Title: Mast cell exosomes promote lung adenocarcinoma cell proliferation – role of KIT-stem cell factor signaling

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/s12964-014-0064-8

    Mast cell-derived exosomes can activate the KIT-SCF signaling pathway in A549 cells. A549 cells treated with exosomes from control cells (HEK 293) or HMC-1 cells were analyzed using Western blot. Phosphorylated and total PI3K, AKT, GSK3β and cyclinD1 were measured by Western blot and to normalize protein loading, samples were also probed for GAPDH. (A) . Relative intensity was calculated as follows for p-PI3K (B) , p-AKT (C) and p-GSK3β (D) ; (phosphorylated protein/GAPDH)/(total protein/GAPDH), and for cyclin D1 (E) ; cyclin D1/GAPDH. All the above data are representative of three independent experiments (n =3). *P
    Figure Legend Snippet: Mast cell-derived exosomes can activate the KIT-SCF signaling pathway in A549 cells. A549 cells treated with exosomes from control cells (HEK 293) or HMC-1 cells were analyzed using Western blot. Phosphorylated and total PI3K, AKT, GSK3β and cyclinD1 were measured by Western blot and to normalize protein loading, samples were also probed for GAPDH. (A) . Relative intensity was calculated as follows for p-PI3K (B) , p-AKT (C) and p-GSK3β (D) ; (phosphorylated protein/GAPDH)/(total protein/GAPDH), and for cyclin D1 (E) ; cyclin D1/GAPDH. All the above data are representative of three independent experiments (n =3). *P

    Techniques Used: Derivative Assay, Western Blot

    18) Product Images from "Periostin enhances adipose-derived stem cell adhesion, migration, and therapeutic efficiency in Apo E deficient mice with hind limb ischemia"

    Article Title: Periostin enhances adipose-derived stem cell adhesion, migration, and therapeutic efficiency in Apo E deficient mice with hind limb ischemia

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-015-0126-x

    Effects of periostin on adhesion and migration of ADSCs. More attached cells were observed in the P-ADSC group compared with those in the ADSC control and siRNA groups a , and quantitative assays indicated that periostin overexpression effectively enhanced the adhesion of ADSCs b . Migration assays of the ADSCs transfected with and without periostin and siRNA groups were conducted using a Boyden chamber c . Data indicate that periostin overexpression improved the migration of ADSCs d . Scale bar = 100 μm. Western blot analysis revealed upregulation and inhibition of the integrin-β1/FAK/Akt/PI3K/eNOS signaling pathway e . Significant differences were observed between the P-ADSC and ADSC groups and the siRNA group f . * P
    Figure Legend Snippet: Effects of periostin on adhesion and migration of ADSCs. More attached cells were observed in the P-ADSC group compared with those in the ADSC control and siRNA groups a , and quantitative assays indicated that periostin overexpression effectively enhanced the adhesion of ADSCs b . Migration assays of the ADSCs transfected with and without periostin and siRNA groups were conducted using a Boyden chamber c . Data indicate that periostin overexpression improved the migration of ADSCs d . Scale bar = 100 μm. Western blot analysis revealed upregulation and inhibition of the integrin-β1/FAK/Akt/PI3K/eNOS signaling pathway e . Significant differences were observed between the P-ADSC and ADSC groups and the siRNA group f . * P

    Techniques Used: Migration, Over Expression, Transfection, Western Blot, Inhibition

    19) Product Images from "HMGA1 participates in MHCC97H cell proliferation and invasion through the ILK/Akt/GSK3β signaling pathway"

    Article Title: HMGA1 participates in MHCC97H cell proliferation and invasion through the ILK/Akt/GSK3β signaling pathway

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.7820

    The effects on cell migration in MHCC97H hepatocellular carcinoma cells treated with shHMGA1, ILK expression vector and/or MK2206. Cell migration was examined using a wound-healing assay (magnification, ×40). DMSO, dimethylsulfoxide; EGFP, enhanced green fluorescent protein; HMGA1, high mobility group AT-hook 1; ILK, integrin-linked kinase; MK2206, an Akt-specific inhibitor; sh, short hairpin RNA.
    Figure Legend Snippet: The effects on cell migration in MHCC97H hepatocellular carcinoma cells treated with shHMGA1, ILK expression vector and/or MK2206. Cell migration was examined using a wound-healing assay (magnification, ×40). DMSO, dimethylsulfoxide; EGFP, enhanced green fluorescent protein; HMGA1, high mobility group AT-hook 1; ILK, integrin-linked kinase; MK2206, an Akt-specific inhibitor; sh, short hairpin RNA.

    Techniques Used: Migration, Expressing, Plasmid Preparation, Wound Healing Assay, shRNA

    The effects on apoptosis in MHCC97H hepatocellular carcinoma cells treated with shHMGA1, ILK expression vector and/or MK2206. In each panel, B2 quadrant indicates only PI positive cells that were necrotic and the B4 quadrant indicates live cells. The B1 quadrant indicates Annexin and PI as late apoptosis cells and the B3 quadrant illustrates Annexin as early apoptotic cells. Apoptosis levels were also detected by flow cytometry in each of the seven experimental groups: (A) Blank control group; (B) shControl group; (C) shHMGA1 group; (D) shHMGA1 + EGFP group; (E) shHMGA1 + ILK group; (F) shHMGA1 + ILK + DMSO group; and (G) shHMGA1 + ILK + MK2206 group. DMSO, dimethylsulfoxide; EGFP, enhanced green fluorescent protein; HMGA1, high mobility group AT-hook 1; ILK, integrin-linked kinase; MK2206, an Akt-specific inhibitor; sh, short hairpin RNA.
    Figure Legend Snippet: The effects on apoptosis in MHCC97H hepatocellular carcinoma cells treated with shHMGA1, ILK expression vector and/or MK2206. In each panel, B2 quadrant indicates only PI positive cells that were necrotic and the B4 quadrant indicates live cells. The B1 quadrant indicates Annexin and PI as late apoptosis cells and the B3 quadrant illustrates Annexin as early apoptotic cells. Apoptosis levels were also detected by flow cytometry in each of the seven experimental groups: (A) Blank control group; (B) shControl group; (C) shHMGA1 group; (D) shHMGA1 + EGFP group; (E) shHMGA1 + ILK group; (F) shHMGA1 + ILK + DMSO group; and (G) shHMGA1 + ILK + MK2206 group. DMSO, dimethylsulfoxide; EGFP, enhanced green fluorescent protein; HMGA1, high mobility group AT-hook 1; ILK, integrin-linked kinase; MK2206, an Akt-specific inhibitor; sh, short hairpin RNA.

    Techniques Used: Expressing, Plasmid Preparation, Flow Cytometry, Cytometry, shRNA

    mRNA and protein expression levels of HMGA1, ILK, p-Akt and p-GSK3β in MHCC97H hepatocellular carcinoma cells treated with shHMGA1, ILK expression vector and/or MK2206. Alterations in mRNA expression levels of (A) HMGA1 and (B) ILK were evaluated by reverse transcription-quantitative polymerase chain reaction. *P
    Figure Legend Snippet: mRNA and protein expression levels of HMGA1, ILK, p-Akt and p-GSK3β in MHCC97H hepatocellular carcinoma cells treated with shHMGA1, ILK expression vector and/or MK2206. Alterations in mRNA expression levels of (A) HMGA1 and (B) ILK were evaluated by reverse transcription-quantitative polymerase chain reaction. *P

    Techniques Used: Expressing, Plasmid Preparation, Real-time Polymerase Chain Reaction

    20) Product Images from "SH2B1 promotes NSCLC cell proliferation through PI3K/Akt/mTOR signaling cascade"

    Article Title: SH2B1 promotes NSCLC cell proliferation through PI3K/Akt/mTOR signaling cascade

    Journal: Cancer Cell International

    doi: 10.1186/s12935-018-0632-x

    SH2B1 regulates NSCLC cell proliferation through Akt/mTOR pathway in vitro. a , b The effect of SH2B1 on Akt/mTOR signaling. a The protein levels of SH2B1, p-Akt, p-mTOR and PTEN in different panels of A549 and H1299 cells were assayed by Western blotting. b The protein levels of Akt/mTOR pathway targets (p-Akt, p-mTOR and PTEN) in different panels of H1299, which were subjected or not subjected to a Akt inhibitor, 10 μm/l deguelin for 24H, were assayed by Western blotting. Experiments were repeated three times and representative pictures are shown for a and b . GAPDH is used as an internal control. c CCK8 assay indicated that deguelin impaired the effect of SH2B1 on cell proliferation. Results are presented as mean ± SD
    Figure Legend Snippet: SH2B1 regulates NSCLC cell proliferation through Akt/mTOR pathway in vitro. a , b The effect of SH2B1 on Akt/mTOR signaling. a The protein levels of SH2B1, p-Akt, p-mTOR and PTEN in different panels of A549 and H1299 cells were assayed by Western blotting. b The protein levels of Akt/mTOR pathway targets (p-Akt, p-mTOR and PTEN) in different panels of H1299, which were subjected or not subjected to a Akt inhibitor, 10 μm/l deguelin for 24H, were assayed by Western blotting. Experiments were repeated three times and representative pictures are shown for a and b . GAPDH is used as an internal control. c CCK8 assay indicated that deguelin impaired the effect of SH2B1 on cell proliferation. Results are presented as mean ± SD

    Techniques Used: In Vitro, Western Blot, CCK-8 Assay

    Clinical relevance of SH2B1 and its targets in NSCLC. a Representative expression levels of SH2B1, p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN in NSCLC by IHC staining. Scale bars, 10 μm. b Percentages of specimens showing high or low SH2B1 expression relative to levels of p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN (all P
    Figure Legend Snippet: Clinical relevance of SH2B1 and its targets in NSCLC. a Representative expression levels of SH2B1, p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN in NSCLC by IHC staining. Scale bars, 10 μm. b Percentages of specimens showing high or low SH2B1 expression relative to levels of p-Akt (Ser 473), p-mTOR (Ser 2448) and PTEN (all P

    Techniques Used: Expressing, Immunohistochemistry, Staining

    21) Product Images from "Effects of isoliquiritigenin on ovarian cancer cells"

    Article Title: Effects of isoliquiritigenin on ovarian cancer cells

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S149295

    Summary of the inhibitory effect of ILQ on PI3K/Akt/mTOR pathway members. Abbreviation: ILQ, isoliquiritigenin.
    Figure Legend Snippet: Summary of the inhibitory effect of ILQ on PI3K/Akt/mTOR pathway members. Abbreviation: ILQ, isoliquiritigenin.

    Techniques Used:

    Isoliquiritigenin regulates the members of PI3K/Akt/mTOR pathway in Ishikawa and ES-2 cells. ILQ treatment significantly reduced the expression level of p-Akt, p-mTOR, P70/S6K, and Cyclin D1 in both Ishikawa and ES-2 cells. * P
    Figure Legend Snippet: Isoliquiritigenin regulates the members of PI3K/Akt/mTOR pathway in Ishikawa and ES-2 cells. ILQ treatment significantly reduced the expression level of p-Akt, p-mTOR, P70/S6K, and Cyclin D1 in both Ishikawa and ES-2 cells. * P

    Techniques Used: Expressing

    Isoliquiritigenin regulates the members of PI3K/Akt/mTOR pathway. ( A ) ILQ treatment significantly reduced the phosphorylated form p-Akt and p-mTOR in both SKOV3 and OVCAR3 cells. ( B ) ILQ treatment significantly reduced the expression level of downstream proteins. ( C ) ILQ treatment significantly reduced the expression level of Wnt3a and p-ERK. * P
    Figure Legend Snippet: Isoliquiritigenin regulates the members of PI3K/Akt/mTOR pathway. ( A ) ILQ treatment significantly reduced the phosphorylated form p-Akt and p-mTOR in both SKOV3 and OVCAR3 cells. ( B ) ILQ treatment significantly reduced the expression level of downstream proteins. ( C ) ILQ treatment significantly reduced the expression level of Wnt3a and p-ERK. * P

    Techniques Used: Expressing

    22) Product Images from "miR-200a regulates Rheb-mediated amelioration of insulin resistance after duodenal–jejunal bypass"

    Article Title: miR-200a regulates Rheb-mediated amelioration of insulin resistance after duodenal–jejunal bypass

    Journal: International Journal of Obesity (2005)

    doi: 10.1038/ijo.2016.60

    The effect of miR-200a on the phosphorylation level of downstream mediators in insulin-resistant BRL cell line. ( a ) Analysis and quantification of mTOR, IRS1/2 and AKT phosphorylation. ( b , c ) Analysis and quantification of Rheb and pS473-AKT-phosphorylation in response to insulin in insulin-resistant cells ( b ) or normal cells ( c ). The data correspond to three independent experiments, each of which performed in duplicate, ±s.e., with n =5. NS, not significant, * P
    Figure Legend Snippet: The effect of miR-200a on the phosphorylation level of downstream mediators in insulin-resistant BRL cell line. ( a ) Analysis and quantification of mTOR, IRS1/2 and AKT phosphorylation. ( b , c ) Analysis and quantification of Rheb and pS473-AKT-phosphorylation in response to insulin in insulin-resistant cells ( b ) or normal cells ( c ). The data correspond to three independent experiments, each of which performed in duplicate, ±s.e., with n =5. NS, not significant, * P

    Techniques Used:

    ( a ) Western blot of Rheb, mTOR, IRS1/2 and AKT expression, and their phosphorylation states in liver of DJB and sham rats at week 8 postoperatively at fed state. ( b , c ) IHC staining of liver showing that Rheb was clearly decreased at week 8 postoperatively. ( d ) mRNA expression of PEPCK and G6Pase in liver in DJB and Sham group at week 8 postoperatively at fed state, and variance between each group is similar. * P
    Figure Legend Snippet: ( a ) Western blot of Rheb, mTOR, IRS1/2 and AKT expression, and their phosphorylation states in liver of DJB and sham rats at week 8 postoperatively at fed state. ( b , c ) IHC staining of liver showing that Rheb was clearly decreased at week 8 postoperatively. ( d ) mRNA expression of PEPCK and G6Pase in liver in DJB and Sham group at week 8 postoperatively at fed state, and variance between each group is similar. * P

    Techniques Used: Western Blot, Expressing, Immunohistochemistry, Staining

    23) Product Images from "Silencing novel long non-coding RNA FKBP9P1 represses malignant progression and inhibits PI3K/AKT signaling of head and neck squamous cell carcinoma in vitro"

    Article Title: Silencing novel long non-coding RNA FKBP9P1 represses malignant progression and inhibits PI3K/AKT signaling of head and neck squamous cell carcinoma in vitro

    Journal: Chinese Medical Journal

    doi: 10.1097/CM9.0000000000000933

    Knockdown of FKBP9P1 suppresses the PI3K/AKT signaling pathway in HNSCC cells. Western blotting results demonstrated that knockdown of FKBP9P1 suppresses the p-PI3K and p-AKT protein expression in HNSCC cells. FKBP9P1: FKBP prolyl isomerase 9 pseudogene 1; HNSCC: Head and neck squamous cell carcinoma; p-PI3K: Phosphorylation of PI3K; p-AKT: Phosphorylation of AKT.
    Figure Legend Snippet: Knockdown of FKBP9P1 suppresses the PI3K/AKT signaling pathway in HNSCC cells. Western blotting results demonstrated that knockdown of FKBP9P1 suppresses the p-PI3K and p-AKT protein expression in HNSCC cells. FKBP9P1: FKBP prolyl isomerase 9 pseudogene 1; HNSCC: Head and neck squamous cell carcinoma; p-PI3K: Phosphorylation of PI3K; p-AKT: Phosphorylation of AKT.

    Techniques Used: Western Blot, Expressing

    24) Product Images from "PTBP1 promotes the growth of breast cancer cells through the PTEN/Akt pathway and autophagy. PTBP1 promotes the growth of breast cancer cells through the PTEN/Akt pathway and autophagy"

    Article Title: PTBP1 promotes the growth of breast cancer cells through the PTEN/Akt pathway and autophagy. PTBP1 promotes the growth of breast cancer cells through the PTEN/Akt pathway and autophagy

    Journal: Journal of Cellular Physiology

    doi: 10.1002/jcp.26823

    Increased expression levels of PTBP1 and p‐Akt were demonstrated in clinical tumor samples from breast cancer patients. (a) PTBP1, p‐Akt, and Akt expressions were analyzed in 137 breast cancer samples as determined by western blot. Actin was used as the control. (b) The relationship between PTBP1 and p‐Akt was significant. P
    Figure Legend Snippet: Increased expression levels of PTBP1 and p‐Akt were demonstrated in clinical tumor samples from breast cancer patients. (a) PTBP1, p‐Akt, and Akt expressions were analyzed in 137 breast cancer samples as determined by western blot. Actin was used as the control. (b) The relationship between PTBP1 and p‐Akt was significant. P

    Techniques Used: Expressing, Western Blot

    Knockdown of PTBP1 leads to upregulation of PTEN and decreased expression of p‐Akt and cell growth. (a) To validate the effct of PTBP1 on P‐Akt western blot analysis was performed. Expression of p‐Akt decreased in PTBP1‐knockdown cells compared with control. GADPH was used as a loading control. (b) With the overexpression of PTBP1, the expression of p‐Akt increased compared with control using western blot assay. GADPH was used as a loading control. (c) To validate the effct of PTBP1 on PTEN, western blot analysis was performed. The expression of PTEN increased in PTBP1‐knockdown cells compared with the control. GADPH was used as a loading control. (d) The overexpression of PTBP1 decreased the expression of PTEN compared with the controls using western blot assay. GADPH was used as a loading control. (e) To validate the effct of PTBP1 on autophagy, the expression level of LC3BI and LC3BII was examined by western blot analysis. Knockdown of PTBP1 induced the transition of the LC3BI to LC3BII. GADPH was used as a loading control. (f) Overexpression of PTBP1 reduced the transition of the LC3BI to LC3BII using western blot analysis. GADPH was used as a loading control. GADPH, glyceraldehyde‐3‐phosphate dehydrogenase; p‐AkPTBP1, polypyrimidine tract binding protein 1; PTEN, phosphatase and tensin homolog
    Figure Legend Snippet: Knockdown of PTBP1 leads to upregulation of PTEN and decreased expression of p‐Akt and cell growth. (a) To validate the effct of PTBP1 on P‐Akt western blot analysis was performed. Expression of p‐Akt decreased in PTBP1‐knockdown cells compared with control. GADPH was used as a loading control. (b) With the overexpression of PTBP1, the expression of p‐Akt increased compared with control using western blot assay. GADPH was used as a loading control. (c) To validate the effct of PTBP1 on PTEN, western blot analysis was performed. The expression of PTEN increased in PTBP1‐knockdown cells compared with the control. GADPH was used as a loading control. (d) The overexpression of PTBP1 decreased the expression of PTEN compared with the controls using western blot assay. GADPH was used as a loading control. (e) To validate the effct of PTBP1 on autophagy, the expression level of LC3BI and LC3BII was examined by western blot analysis. Knockdown of PTBP1 induced the transition of the LC3BI to LC3BII. GADPH was used as a loading control. (f) Overexpression of PTBP1 reduced the transition of the LC3BI to LC3BII using western blot analysis. GADPH was used as a loading control. GADPH, glyceraldehyde‐3‐phosphate dehydrogenase; p‐AkPTBP1, polypyrimidine tract binding protein 1; PTEN, phosphatase and tensin homolog

    Techniques Used: Expressing, Western Blot, Over Expression, Binding Assay

    25) Product Images from "Application of citrate as a tricarboxylic acid (TCA) cycle intermediate, prevents diabetic-induced heart damages in mice"

    Article Title: Application of citrate as a tricarboxylic acid (TCA) cycle intermediate, prevents diabetic-induced heart damages in mice

    Journal: Iranian Journal of Basic Medical Sciences

    doi:

    Citrate application prevents diabetes-induced accumulation of pro-inflammatory and apoptotic cytokines in the heart. Caspase-3, plasminogen activator inhibitor 1 (PAI1), p-AKT and AKT levels were assessed by Western blot analysis in cardiac tissues of non-diabetic and diabetic mice with or without citrate application after 8 weeks. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as a loading control and the black arrowhead on the right side of the blot indicates Caspase-3 band (A). Semi-quantitative analysis of Caspase-3, PAI1, p-AKT and AKT normalized to GAPDH (B). Data are presented as mean±SEM. Each group included six mice, significant differences between non-DM and DM or DM and DM+Citrate groups were shown (* P -value
    Figure Legend Snippet: Citrate application prevents diabetes-induced accumulation of pro-inflammatory and apoptotic cytokines in the heart. Caspase-3, plasminogen activator inhibitor 1 (PAI1), p-AKT and AKT levels were assessed by Western blot analysis in cardiac tissues of non-diabetic and diabetic mice with or without citrate application after 8 weeks. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as a loading control and the black arrowhead on the right side of the blot indicates Caspase-3 band (A). Semi-quantitative analysis of Caspase-3, PAI1, p-AKT and AKT normalized to GAPDH (B). Data are presented as mean±SEM. Each group included six mice, significant differences between non-DM and DM or DM and DM+Citrate groups were shown (* P -value

    Techniques Used: Western Blot, Mouse Assay

    26) Product Images from "miR-185 suppresses progression of Ewing’s sarcoma via inhibiting the PI3K/AKT and Wnt/β-catenin pathways"

    Article Title: miR-185 suppresses progression of Ewing’s sarcoma via inhibiting the PI3K/AKT and Wnt/β-catenin pathways

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S167771

    Overexpression of miR-185 suppresses the PI3K/Akt/mTOR and Wnt/β-catenin pathways in RD-ES cells. Notes: After transfection for 48 hours, Western blot assays of PI3K/Akt/mTOR pathway-related proteins ( A ) and Wnt/β-catenin pathway-related proteins ( B ) were conducted in RD-ES cells. Levels of proteins were normalized to GAPDH, and the relative expression of the corresponding protein in miR-185 overexpressed cells was normalized to the NC. ( C ) After transfection for 48 hours, immunohistochemistry assay was performed to detect the expression of β-catenin and E-cad. The magnification was ×400. Data are presented as the mean ± standard deviation (n=3). Results were obtained in three replicates. miR-185, transfected with pCMV-MIR-miR-185 vector; NC, negative control, transfected with pCMV-MIR empty vector. * P
    Figure Legend Snippet: Overexpression of miR-185 suppresses the PI3K/Akt/mTOR and Wnt/β-catenin pathways in RD-ES cells. Notes: After transfection for 48 hours, Western blot assays of PI3K/Akt/mTOR pathway-related proteins ( A ) and Wnt/β-catenin pathway-related proteins ( B ) were conducted in RD-ES cells. Levels of proteins were normalized to GAPDH, and the relative expression of the corresponding protein in miR-185 overexpressed cells was normalized to the NC. ( C ) After transfection for 48 hours, immunohistochemistry assay was performed to detect the expression of β-catenin and E-cad. The magnification was ×400. Data are presented as the mean ± standard deviation (n=3). Results were obtained in three replicates. miR-185, transfected with pCMV-MIR-miR-185 vector; NC, negative control, transfected with pCMV-MIR empty vector. * P

    Techniques Used: Over Expression, Transfection, Western Blot, Expressing, Immunohistochemistry, Standard Deviation, Plasmid Preparation, Negative Control

    27) Product Images from "Pterostilbene increases PTEN expression through the targeted downregulation of microRNA-19a in hepatocellular carcinoma"

    Article Title: Pterostilbene increases PTEN expression through the targeted downregulation of microRNA-19a in hepatocellular carcinoma

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2018.8515

    PTEN, Akt and p-Akt levels in SMMC-7721 cells following miR-19a inhibitor transfection and Pter treatment for 24 h. (A) PTEN, Akt and p-Akt levels in SMMC-7721 cells were detected by western blot analysis and representative bands are presented. (B) Relative quantification of western blot analysis results by ImageJ analysis software. (C) Schematic representation of the proposed model of Pter mediation of the PTEN/Akt signaling pathway via miR-19a. **P
    Figure Legend Snippet: PTEN, Akt and p-Akt levels in SMMC-7721 cells following miR-19a inhibitor transfection and Pter treatment for 24 h. (A) PTEN, Akt and p-Akt levels in SMMC-7721 cells were detected by western blot analysis and representative bands are presented. (B) Relative quantification of western blot analysis results by ImageJ analysis software. (C) Schematic representation of the proposed model of Pter mediation of the PTEN/Akt signaling pathway via miR-19a. **P

    Techniques Used: Transfection, Western Blot, Software

    28) Product Images from "Trastuzumab induces PUMA‐dependent apoptosis and inhibits tumor growth in gastric cancer"

    Article Title: Trastuzumab induces PUMA‐dependent apoptosis and inhibits tumor growth in gastric cancer

    Journal: FEBS Open Bio

    doi: 10.1002/2211-5463.12522

    The PUMA induction by trastuzumab is mediated through GSK 3β activation. (A) NCI ‐N87 cells were transfected with either a control scrambled si RNA or a GSK 3 β si RNA for 24 h and then treated with 10 μmol·L −1 trastuzumab for 6 h. Nuclear fractions were isolated from cells treated with trastuzumab and analyzed for p65 and GSK 3β expression by western blotting. (B) NCI ‐N87 cells were transfected with either a control scrambled si RNA or a GSK 3 β si RNA for 24 h and then treated with 10 μmol·L −1 trastuzumab for 24 h. GSK 3β and PUMA expression was analyzed by western blotting. (C) NCI ‐N87 cells were treated with 10 μmol·L −1 trastuzumab at indicated time point. Relative protein expression was analyzed by western blotting. (D) NCI ‐N87 cells were transfected with active AKT plasmid for 8 h and then treated with 10 μmol·L −1 trastuzumab for 24 h. PUMA , p‐ AKT , and total AKT expression was analyzed by western blotting.
    Figure Legend Snippet: The PUMA induction by trastuzumab is mediated through GSK 3β activation. (A) NCI ‐N87 cells were transfected with either a control scrambled si RNA or a GSK 3 β si RNA for 24 h and then treated with 10 μmol·L −1 trastuzumab for 6 h. Nuclear fractions were isolated from cells treated with trastuzumab and analyzed for p65 and GSK 3β expression by western blotting. (B) NCI ‐N87 cells were transfected with either a control scrambled si RNA or a GSK 3 β si RNA for 24 h and then treated with 10 μmol·L −1 trastuzumab for 24 h. GSK 3β and PUMA expression was analyzed by western blotting. (C) NCI ‐N87 cells were treated with 10 μmol·L −1 trastuzumab at indicated time point. Relative protein expression was analyzed by western blotting. (D) NCI ‐N87 cells were transfected with active AKT plasmid for 8 h and then treated with 10 μmol·L −1 trastuzumab for 24 h. PUMA , p‐ AKT , and total AKT expression was analyzed by western blotting.

    Techniques Used: Activation Assay, Transfection, Isolation, Expressing, Western Blot, Plasmid Preparation

    29) Product Images from "Achyranthes bidentata Polypeptide Protects Schwann Cells From Apoptosis in Hydrogen Peroxide-Induced Oxidative Stress"

    Article Title: Achyranthes bidentata Polypeptide Protects Schwann Cells From Apoptosis in Hydrogen Peroxide-Induced Oxidative Stress

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2018.00868

    ABPPk treatment activates the PI3K/AKT and ERK1/2 pathways in SCs. (A) SCs were treated with 400 μM H 2 O 2 with or without 0.5 μg/ml ABPPk. The cells were collected and analyzed by Western blot for the expression of p-PI3K and PI3K, p-AKT and AKT, and p-ERK1/2 and ERK1/2. (B) Histogram shows data of p-PI3K/PI3K, p-AKT/AKT, and p-ERK/ERK levels from Western blot analyses. GAPDH was used as the protein loading control and band density normalization. ABPPk vs. control: P
    Figure Legend Snippet: ABPPk treatment activates the PI3K/AKT and ERK1/2 pathways in SCs. (A) SCs were treated with 400 μM H 2 O 2 with or without 0.5 μg/ml ABPPk. The cells were collected and analyzed by Western blot for the expression of p-PI3K and PI3K, p-AKT and AKT, and p-ERK1/2 and ERK1/2. (B) Histogram shows data of p-PI3K/PI3K, p-AKT/AKT, and p-ERK/ERK levels from Western blot analyses. GAPDH was used as the protein loading control and band density normalization. ABPPk vs. control: P

    Techniques Used: Western Blot, Expressing

    Effects of the LY294002 and PD98059, PI3K inhibitor, and ERK1/2 inhibitor, on oxidative damage attenuated by ABPPk in SCs. (A) Western blot shows the p-PI3K/PI3K, p-AKT/AKT, and p-ERK/ERK in SCs pretreated with LY294002 (10 μM) and PD98059 (10 μM) for 30 min before the treatment of H 2 O 2 (400 μM) and ABPPk (0.5 μg/ml) for 24 h. (B) Densitometric analyses illustrates the results of p-PI3K/PI3K, p-AKT/AKT, and p-ERK/ERK. H 2 O 2 vs control: ∗ P
    Figure Legend Snippet: Effects of the LY294002 and PD98059, PI3K inhibitor, and ERK1/2 inhibitor, on oxidative damage attenuated by ABPPk in SCs. (A) Western blot shows the p-PI3K/PI3K, p-AKT/AKT, and p-ERK/ERK in SCs pretreated with LY294002 (10 μM) and PD98059 (10 μM) for 30 min before the treatment of H 2 O 2 (400 μM) and ABPPk (0.5 μg/ml) for 24 h. (B) Densitometric analyses illustrates the results of p-PI3K/PI3K, p-AKT/AKT, and p-ERK/ERK. H 2 O 2 vs control: ∗ P

    Techniques Used: Western Blot

    The protective ABPPk on apoptosis induced by H 2 O 2 are partly impaired through the inhibition of the PI3K/AKT and ERK1/2 pathways. (A) Protein expression levels of cleaved caspase-3. (B) Quantification data of apoptotic markers cleaved caspase-3 in each group. H 2 O 2 vs control: ∗ P
    Figure Legend Snippet: The protective ABPPk on apoptosis induced by H 2 O 2 are partly impaired through the inhibition of the PI3K/AKT and ERK1/2 pathways. (A) Protein expression levels of cleaved caspase-3. (B) Quantification data of apoptotic markers cleaved caspase-3 in each group. H 2 O 2 vs control: ∗ P

    Techniques Used: Inhibition, Expressing

    30) Product Images from "Luteolin attenuates doxorubicin-induced cardiotoxicity by modulating the PHLPP1/AKT/Bcl-2 signalling pathway"

    Article Title: Luteolin attenuates doxorubicin-induced cardiotoxicity by modulating the PHLPP1/AKT/Bcl-2 signalling pathway

    Journal: PeerJ

    doi: 10.7717/peerj.8845

    Chemicals-target gene network linking the protective effects of LUT against cardiotoxicity to potential signalling pathways and related protein expression. (A) Blue circles represent the enriched KEGG main pathway, and green rectangles represent the top putative target proteins. (B–D) Representative phlpp1 and GAPDH protein ratio. (E–G) p-AKT and AKT protein ratio. ∗ p
    Figure Legend Snippet: Chemicals-target gene network linking the protective effects of LUT against cardiotoxicity to potential signalling pathways and related protein expression. (A) Blue circles represent the enriched KEGG main pathway, and green rectangles represent the top putative target proteins. (B–D) Representative phlpp1 and GAPDH protein ratio. (E–G) p-AKT and AKT protein ratio. ∗ p

    Techniques Used: Expressing

    31) Product Images from "Effects of Arf6 downregulation on biological characteristics of human prostate cancer cells"

    Article Title: Effects of Arf6 downregulation on biological characteristics of human prostate cancer cells

    Journal: International Brazilian Journal of Urology : official journal of the Brazilian Society of Urology

    doi: 10.1590/S1677-5538.IBJU.2019.0499

    Expressions of key signaling molecules in PI3K/AKT and ERK/Rac1 pathways detected by Western blot. Con: Normal control group, NC: negative control group, siRNA: siRNA interference group. **Compared with Con and NC groups, P
    Figure Legend Snippet: Expressions of key signaling molecules in PI3K/AKT and ERK/Rac1 pathways detected by Western blot. Con: Normal control group, NC: negative control group, siRNA: siRNA interference group. **Compared with Con and NC groups, P

    Techniques Used: Western Blot, Negative Control

    32) Product Images from "Tanshinone IIA Reverses Gefitinib-Resistance In Human Non-Small-Cell Lung Cancer Via Regulation Of VEGFR/Akt Pathway"

    Article Title: Tanshinone IIA Reverses Gefitinib-Resistance In Human Non-Small-Cell Lung Cancer Via Regulation Of VEGFR/Akt Pathway

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S221228

    Tan IIA enhances the sensitivity of HCC827/gefitinib cells to gefitinib by regulation of VEGFR/Akt pathway. HCC827/gefitinib cells were treated with 40 nM OXA or/and 2 μM Tan IIA, or OXA + Tan IIA + VEGF for 72 hrs. ( A ) Expression levels of p-EGFR, p-VEGFR2 and p-Akt in HCC827/gefitinib cells were detected with Western blotting. β-actin was used as an internal control. ( B–D ) The relative expressions of p-EGFR, p-VEGFR2 and p-Akt in HCC827/gefitinib cells were quantified via normalization to β-actin. *P
    Figure Legend Snippet: Tan IIA enhances the sensitivity of HCC827/gefitinib cells to gefitinib by regulation of VEGFR/Akt pathway. HCC827/gefitinib cells were treated with 40 nM OXA or/and 2 μM Tan IIA, or OXA + Tan IIA + VEGF for 72 hrs. ( A ) Expression levels of p-EGFR, p-VEGFR2 and p-Akt in HCC827/gefitinib cells were detected with Western blotting. β-actin was used as an internal control. ( B–D ) The relative expressions of p-EGFR, p-VEGFR2 and p-Akt in HCC827/gefitinib cells were quantified via normalization to β-actin. *P

    Techniques Used: Expressing, Western Blot

    Tan IIA enhances the sensitivity of HCC827/gefitinib cells to gefitinib in vivo. HCC827/gefitinib cells were injected subcutaneously into nude mice to form subcutaneous tumors. ( A ) Tumor volumes of mice were measured weekly. ( B ) HCC827/gefitinib xenograft tumors were excised from xenografts and pictured on day 21. ( C ) The weight in each group of mice was calculated on day 21. ( D ) Expression levels of p-EGFR, p-VEGFR2 and p-Akt in tumor tissues were measured with Western blotting. ( E, F, G ) The relative expressions of p-EGFR, p-VEGFR2 and p-Akt were quantified via normalization to β-actin. *P
    Figure Legend Snippet: Tan IIA enhances the sensitivity of HCC827/gefitinib cells to gefitinib in vivo. HCC827/gefitinib cells were injected subcutaneously into nude mice to form subcutaneous tumors. ( A ) Tumor volumes of mice were measured weekly. ( B ) HCC827/gefitinib xenograft tumors were excised from xenografts and pictured on day 21. ( C ) The weight in each group of mice was calculated on day 21. ( D ) Expression levels of p-EGFR, p-VEGFR2 and p-Akt in tumor tissues were measured with Western blotting. ( E, F, G ) The relative expressions of p-EGFR, p-VEGFR2 and p-Akt were quantified via normalization to β-actin. *P

    Techniques Used: In Vivo, Injection, Mouse Assay, Expressing, Western Blot

    33) Product Images from "Epothilone B Facilitates Peripheral Nerve Regeneration by Promoting Autophagy and Migration in Schwann Cells"

    Article Title: Epothilone B Facilitates Peripheral Nerve Regeneration by Promoting Autophagy and Migration in Schwann Cells

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2020.00143

    EpoB significantly enhances migration by PI3K/Akt signaling-mediated autophagy in SCs. Representative western blotting (A) and data analysis of p-PI3K and p-Akt in each group (B) . (C) Quantification of the total number of autophagosomes in each group. Transwell analysis of migratory cells in the control (D) , EpoB (E) , EpoB+IGF-1 (F) , LY294002 (G) , and LY294002+3-MA (H) groups at 24 h after treatment. (I) Quantification of the total number of migratory cells in each group. TEM analysis showing the formation of autophagosomes in SCs from the control (J,N) , EpoB (K,O) , EpoB+IGF-1 (L,P) , and EpoB+ LY294002 (M,Q) groups at 24 h after treatment. White arrowheads indicate normal endoplasmic reticula; black arrowheads indicate double-membraned autophagosomes. Scale bars: (J–M) 4 μm; (N–Q) 1 μm. * P
    Figure Legend Snippet: EpoB significantly enhances migration by PI3K/Akt signaling-mediated autophagy in SCs. Representative western blotting (A) and data analysis of p-PI3K and p-Akt in each group (B) . (C) Quantification of the total number of autophagosomes in each group. Transwell analysis of migratory cells in the control (D) , EpoB (E) , EpoB+IGF-1 (F) , LY294002 (G) , and LY294002+3-MA (H) groups at 24 h after treatment. (I) Quantification of the total number of migratory cells in each group. TEM analysis showing the formation of autophagosomes in SCs from the control (J,N) , EpoB (K,O) , EpoB+IGF-1 (L,P) , and EpoB+ LY294002 (M,Q) groups at 24 h after treatment. White arrowheads indicate normal endoplasmic reticula; black arrowheads indicate double-membraned autophagosomes. Scale bars: (J–M) 4 μm; (N–Q) 1 μm. * P

    Techniques Used: Migration, Western Blot, Transmission Electron Microscopy

    34) Product Images from "MiR-25-3p promotes the proliferation of triple negative breast cancer by targeting BTG2"

    Article Title: MiR-25-3p promotes the proliferation of triple negative breast cancer by targeting BTG2

    Journal: Molecular Cancer

    doi: 10.1186/s12943-017-0754-0

    miR-25-3p functioned through BTG2 in AKT and ERK MAPK signaling pathway. a , b . western blot for BTG2, AKT, p-AKT, ERK1/2 and p-ERK1/2 in MDA-MB-231 and Sum-1315 cells transfected with inhibitor-control (inhibitor-ctrl) and siBTG2 scrambled oligonucleotide (si-ctrl), inhibitor and si-ctrl, inhibitor and siBTG2, or inhibitor-ctrl and siBTG2. Protein expression was quantified by band intensity and normalized to GAPDH. * p
    Figure Legend Snippet: miR-25-3p functioned through BTG2 in AKT and ERK MAPK signaling pathway. a , b . western blot for BTG2, AKT, p-AKT, ERK1/2 and p-ERK1/2 in MDA-MB-231 and Sum-1315 cells transfected with inhibitor-control (inhibitor-ctrl) and siBTG2 scrambled oligonucleotide (si-ctrl), inhibitor and si-ctrl, inhibitor and siBTG2, or inhibitor-ctrl and siBTG2. Protein expression was quantified by band intensity and normalized to GAPDH. * p

    Techniques Used: Western Blot, Multiple Displacement Amplification, Transfection, Expressing

    35) Product Images from "Apigenin suppresses the apoptosis of H9C2 rat cardiomyocytes subjected to myocardial ischemia-reperfusion injury via upregulation of the PI3K/Akt pathway"

    Article Title: Apigenin suppresses the apoptosis of H9C2 rat cardiomyocytes subjected to myocardial ischemia-reperfusion injury via upregulation of the PI3K/Akt pathway

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2018.9115

    Apigenin affects the PI3K/Akt pathway. H9C2 cells were treated with MI/R, and 1, 6 and 25 µM apigenin + MI/R. Western blot analysis was conducted to measure the expression levels of (A) p-PI3K and PI3K and (B) p-Akt and Akt in H9C2 cells. *P
    Figure Legend Snippet: Apigenin affects the PI3K/Akt pathway. H9C2 cells were treated with MI/R, and 1, 6 and 25 µM apigenin + MI/R. Western blot analysis was conducted to measure the expression levels of (A) p-PI3K and PI3K and (B) p-Akt and Akt in H9C2 cells. *P

    Techniques Used: Western Blot, Expressing

    36) Product Images from "Astragalus polysaccharide alleviates LPS-induced inflammation injury by regulating miR-127 in H9c2 cardiomyoblasts"

    Article Title: Astragalus polysaccharide alleviates LPS-induced inflammation injury by regulating miR-127 in H9c2 cardiomyoblasts

    Journal: International Journal of Immunopathology and Pharmacology

    doi: 10.1177/2058738418759180

    APS reduced LPS-induced inflammation injury by down-regulating miR-127 and inhibiting NF-κB and JNK and promoting PI3K/AKT signaling pathways in H9c2 cells. (a) H9c2 cells were treated with LPS or co-treated with APS and LPS, and then expression levels of miR-127 in H9c2 cells were measured by qRT-PCR. (b)–(d) Protein expression levels of p65/p-p65, lκBα/p-lκBα JNK/p-JNK, c-Jun/p-c-Jun, PI3K/p-PI3K, and AKT/p-AKT were detected by western blot. Different letters above the bars (a, b, c) indicate that the means of different groups were significantly different ( P
    Figure Legend Snippet: APS reduced LPS-induced inflammation injury by down-regulating miR-127 and inhibiting NF-κB and JNK and promoting PI3K/AKT signaling pathways in H9c2 cells. (a) H9c2 cells were treated with LPS or co-treated with APS and LPS, and then expression levels of miR-127 in H9c2 cells were measured by qRT-PCR. (b)–(d) Protein expression levels of p65/p-p65, lκBα/p-lκBα JNK/p-JNK, c-Jun/p-c-Jun, PI3K/p-PI3K, and AKT/p-AKT were detected by western blot. Different letters above the bars (a, b, c) indicate that the means of different groups were significantly different ( P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    37) Product Images from "Astragalus polysaccharide alleviates LPS-induced inflammation injury by regulating miR-127 in H9c2 cardiomyoblasts"

    Article Title: Astragalus polysaccharide alleviates LPS-induced inflammation injury by regulating miR-127 in H9c2 cardiomyoblasts

    Journal: International Journal of Immunopathology and Pharmacology

    doi: 10.1177/2058738418759180

    APS reduced LPS-induced inflammation injury by down-regulating miR-127 and inhibiting NF-κB and JNK and promoting PI3K/AKT signaling pathways in H9c2 cells. (a) H9c2 cells were treated with LPS or co-treated with APS and LPS, and then expression levels of miR-127 in H9c2 cells were measured by qRT-PCR. (b)–(d) Protein expression levels of p65/p-p65, lκBα/p-lκBα JNK/p-JNK, c-Jun/p-c-Jun, PI3K/p-PI3K, and AKT/p-AKT were detected by western blot. Different letters above the bars (a, b, c) indicate that the means of different groups were significantly different ( P
    Figure Legend Snippet: APS reduced LPS-induced inflammation injury by down-regulating miR-127 and inhibiting NF-κB and JNK and promoting PI3K/AKT signaling pathways in H9c2 cells. (a) H9c2 cells were treated with LPS or co-treated with APS and LPS, and then expression levels of miR-127 in H9c2 cells were measured by qRT-PCR. (b)–(d) Protein expression levels of p65/p-p65, lκBα/p-lκBα JNK/p-JNK, c-Jun/p-c-Jun, PI3K/p-PI3K, and AKT/p-AKT were detected by western blot. Different letters above the bars (a, b, c) indicate that the means of different groups were significantly different ( P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    38) Product Images from "Cytisine, a Partial Agonist of α4β2 Nicotinic Acetylcholine Receptors, Reduced Unpredictable Chronic Mild Stress-Induced Depression-Like Behaviors"

    Article Title: Cytisine, a Partial Agonist of α4β2 Nicotinic Acetylcholine Receptors, Reduced Unpredictable Chronic Mild Stress-Induced Depression-Like Behaviors

    Journal: Biomolecules & Therapeutics

    doi: 10.4062/biomolther.2015.113

    CYT increased mTOR signaling activities in the amygdala. (A) Representative Western blot of proteins in amygdala. (B) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-AKT/AKT in UCMS mice. (C) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-mTOR/mTOR in UCMS mice. (D) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-S6K/S6K in UCMS mice. (E) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-CREB/CREB in UCMS mice. n=6/group. * p
    Figure Legend Snippet: CYT increased mTOR signaling activities in the amygdala. (A) Representative Western blot of proteins in amygdala. (B) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-AKT/AKT in UCMS mice. (C) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-mTOR/mTOR in UCMS mice. (D) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-S6K/S6K in UCMS mice. (E) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-CREB/CREB in UCMS mice. n=6/group. * p

    Techniques Used: Western Blot, Mouse Assay

    CYT increased mTOR signaling activities in the hippocampus. (A) Representative Western blot of proteins in hippocampus. (B) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-AKT/AKT in UCMS mice. (C) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-mTOR/mTOR in UCMS mice. (D) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-S6K/S6K in UCMS mice. (E) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-CREB/CREB in UCMS mice. n=6/group. * p
    Figure Legend Snippet: CYT increased mTOR signaling activities in the hippocampus. (A) Representative Western blot of proteins in hippocampus. (B) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-AKT/AKT in UCMS mice. (C) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-mTOR/mTOR in UCMS mice. (D) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-S6K/S6K in UCMS mice. (E) Treatment of CYT (1 mg/kg) up-regulated the ratio of p-CREB/CREB in UCMS mice. n=6/group. * p

    Techniques Used: Western Blot, Mouse Assay

    39) Product Images from "Matrine attenuates high‐fat diet‐induced in vivo and ox‐LDL‐induced in vitro vascular injury by regulating the PKCα/eNOS and PI3K/Akt/eNOS pathways, et al. Matrine attenuates high‐fat diet‐induced in vivo and ox‐LDL‐induced in vitro vascular injury by regulating the PKCα/eNOS and PI3K/Akt/eNOS pathways"

    Article Title: Matrine attenuates high‐fat diet‐induced in vivo and ox‐LDL‐induced in vitro vascular injury by regulating the PKCα/eNOS and PI3K/Akt/eNOS pathways, et al. Matrine attenuates high‐fat diet‐induced in vivo and ox‐LDL‐induced in vitro vascular injury by regulating the PKCα/eNOS and PI3K/Akt/eNOS pathways

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.14180

    Effects of matrine on eNOS activity and PI3K/Akt/eNOS pathway‐related protein expression in ox‐LDL exposed HUVECs. A, Changes in eNOS activity in HUVECs exposed to ox‐LDL with or without matrine pretreatment along with matrine and/or LY‐294002/L‐NAME as measured using an eNOS activity assay kit. B‐E, Phosphorylation levels of Ser473Akt, Ser1177eNOS, Thr495eNOS, total Akt and the eNOS protein were measured by Western blotting. Representative images of three experiments; densitometric analysis of phosphorylated proteins was normalized to that of total proteins. Data are expressed as the mean ± SD of three independent experiments, * P
    Figure Legend Snippet: Effects of matrine on eNOS activity and PI3K/Akt/eNOS pathway‐related protein expression in ox‐LDL exposed HUVECs. A, Changes in eNOS activity in HUVECs exposed to ox‐LDL with or without matrine pretreatment along with matrine and/or LY‐294002/L‐NAME as measured using an eNOS activity assay kit. B‐E, Phosphorylation levels of Ser473Akt, Ser1177eNOS, Thr495eNOS, total Akt and the eNOS protein were measured by Western blotting. Representative images of three experiments; densitometric analysis of phosphorylated proteins was normalized to that of total proteins. Data are expressed as the mean ± SD of three independent experiments, * P

    Techniques Used: Activity Assay, Expressing, Western Blot

    40) Product Images from "Hematopoietic-substrate-1 associated protein X-1 (HAX-1) regulates liver cancer cells growth, metastasis, and angiogenesis through Akt"

    Article Title: Hematopoietic-substrate-1 associated protein X-1 (HAX-1) regulates liver cancer cells growth, metastasis, and angiogenesis through Akt

    Journal: Cancer Biology & Therapy

    doi: 10.1080/15384047.2019.1617562

    The effects of silencing or overexpressing hematopoietic-substrate-1 associated protein X-1 (HAX-1) on signaling pathways. (a–l) Western blot was applied to test JAK/STAT3 and Akt signaling pathways-related protein levels in different liver cancer cells.
    Figure Legend Snippet: The effects of silencing or overexpressing hematopoietic-substrate-1 associated protein X-1 (HAX-1) on signaling pathways. (a–l) Western blot was applied to test JAK/STAT3 and Akt signaling pathways-related protein levels in different liver cancer cells.

    Techniques Used: Western Blot

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    Blocking Assay:

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    Software:

    Article Title: ARID1A gene silencing reduces the sensitivity of ovarian clear cell carcinoma to cisplatin
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  • 93
    Abcam anti akt1 phospho s129 antibody epr6150
    Effect of non-toxic doses of CK2 inhibitors on GN11 cell migration. ( A ) GN11 cell viability was tested by MTT assay after 24-h treatment with different doses of TBB or CX-4945 inhibitors and expressed as a percentage of the control (Ctrl) ( n = 3). ( B ) Endogenous CK2 activity ( n = 3) in GN11 cells treated with non-toxic doses of inhibitors (10 µM CX-4945 and 75 µM TBB). Both inhibitors strongly prevented the phosphorylation of CK2-specific target <t>Akt1-S129.</t> ( C ) Scratch assay of GN11 cells treated with 10 µM CX-4945, 75 µM TBB, or vehicle for 24 h ( n = 3). Representative images for each condition are shown on the left. Quantification of cell migration is reported on the right as the area covered by cells after 3 or 7 h. ( D ) Boyden chamber assay of GN11 cells pre-treated with 10 µM CX-4945, 75 µM TBB, or vehicle for 30 min ( n = 3). Representative images for each condition are shown on the left. Quantification of migrated cells is represented as the relative migration expressed in percentage; WT cells were used as the reference. Data are shown as mean ± SEM.; * p
    Anti Akt1 Phospho S129 Antibody Epr6150, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti akt1 phospho s129 antibody epr6150/product/Abcam
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    99
    Abcam anti akt
    <t>ARID1A</t> siRNA increases the expression levels of <t>AKT.</t> Western blotting was performed to quantify the expression levels of AKT. β-actin was used as a control for the expression levels. Data are presented as the mean + standard deviation (n=3). *P
    Anti Akt, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti akt/product/Abcam
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    Abcam anti t protein kinase 3 akt antibody
    Kaempferol inactivates <t>PI3K/AKT/mTOR</t> signaling pathway in HepG2 cells by down-regulating miR-21. (a) Relative expressions of PTEN in HepG2 cells after 50 μM kaempferol treatment and/or miR-21 mimic transfection were assessed using quantitative reverse transcription PCR (qRT-PCR). (b) Western blotting was conducted to analyze the expressions of t-PI3K, p-PI3K, t-AKT, p-AKT, t-mTOR, p-mTOR, t-S6 K, and p-S6 K in HepG2 cells after 50 μM kaempferol treatment and/or miR-21 mimic transfection. miR-21: microRNA-21; PTEN: phosphatase and tensin homologue; NC: negative control; PI3K: phosphatidylinositol 3-kinase; AKT: protein kinase 3; mTOR: mechanistic target of rapamycin. * P
    Anti T Protein Kinase 3 Akt Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Abcam phosphorylated akt ser 473 antibodies
    Time-dependent activation of <t>Akt</t> and eNOS paralleling PTEN phosphorylation by GTN. Representative Western blots showing PTEN phosphorylation (Ser 380), eNOS phosphorylation (Ser 1177 or 1179), and Akt phosphorylation (Ser <t>473)</t> in (A) BAEC and (B) HMEC treated with vehicle or 500 nM GTN for the indicated amounts of time. Vehicle was added for 10 min in (A) and 15 min in (B). Results show rapid and sustained eNOS phosphorylation at the activation site Ser 1177, Akt phosphorylation at the activation site Ser 473, and PTEN phosphorylation at the inhibitory site Ser 380 by 500 nM GTN. Band intensities for phosphorylated eNOS in BAEC and for the experiments performed with HMEC were quantified using ImageJ and the values are reported as density units relative to control; * P
    Phosphorylated Akt Ser 473 Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effect of non-toxic doses of CK2 inhibitors on GN11 cell migration. ( A ) GN11 cell viability was tested by MTT assay after 24-h treatment with different doses of TBB or CX-4945 inhibitors and expressed as a percentage of the control (Ctrl) ( n = 3). ( B ) Endogenous CK2 activity ( n = 3) in GN11 cells treated with non-toxic doses of inhibitors (10 µM CX-4945 and 75 µM TBB). Both inhibitors strongly prevented the phosphorylation of CK2-specific target Akt1-S129. ( C ) Scratch assay of GN11 cells treated with 10 µM CX-4945, 75 µM TBB, or vehicle for 24 h ( n = 3). Representative images for each condition are shown on the left. Quantification of cell migration is reported on the right as the area covered by cells after 3 or 7 h. ( D ) Boyden chamber assay of GN11 cells pre-treated with 10 µM CX-4945, 75 µM TBB, or vehicle for 30 min ( n = 3). Representative images for each condition are shown on the left. Quantification of migrated cells is represented as the relative migration expressed in percentage; WT cells were used as the reference. Data are shown as mean ± SEM.; * p

    Journal: International Journal of Molecular Sciences

    Article Title: Protein Kinase CK2 Subunits Differentially Perturb the Adhesion and Migration of GN11 Cells: A Model of Immature Migrating Neurons

    doi: 10.3390/ijms20235951

    Figure Lengend Snippet: Effect of non-toxic doses of CK2 inhibitors on GN11 cell migration. ( A ) GN11 cell viability was tested by MTT assay after 24-h treatment with different doses of TBB or CX-4945 inhibitors and expressed as a percentage of the control (Ctrl) ( n = 3). ( B ) Endogenous CK2 activity ( n = 3) in GN11 cells treated with non-toxic doses of inhibitors (10 µM CX-4945 and 75 µM TBB). Both inhibitors strongly prevented the phosphorylation of CK2-specific target Akt1-S129. ( C ) Scratch assay of GN11 cells treated with 10 µM CX-4945, 75 µM TBB, or vehicle for 24 h ( n = 3). Representative images for each condition are shown on the left. Quantification of cell migration is reported on the right as the area covered by cells after 3 or 7 h. ( D ) Boyden chamber assay of GN11 cells pre-treated with 10 µM CX-4945, 75 µM TBB, or vehicle for 30 min ( n = 3). Representative images for each condition are shown on the left. Quantification of migrated cells is represented as the relative migration expressed in percentage; WT cells were used as the reference. Data are shown as mean ± SEM.; * p

    Article Snippet: Anti-CK2β (ab76025), anti-phospho-Akt1 S129 antibodies (ab133458), and anti-phospho-CDC37 S13 (ab108360) were from Abcam (Cambridge, UK).

    Techniques: Migration, MTT Assay, Activity Assay, Wound Healing Assay, Boyden Chamber Assay

    ARID1A siRNA increases the expression levels of AKT. Western blotting was performed to quantify the expression levels of AKT. β-actin was used as a control for the expression levels. Data are presented as the mean + standard deviation (n=3). *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: ARID1A gene silencing reduces the sensitivity of ovarian clear cell carcinoma to cisplatin

    doi: 10.3892/etm.2016.3863

    Figure Lengend Snippet: ARID1A siRNA increases the expression levels of AKT. Western blotting was performed to quantify the expression levels of AKT. β-actin was used as a control for the expression levels. Data are presented as the mean + standard deviation (n=3). *P

    Article Snippet: The membranes were blocked using 5% skimmed milk powder at room temperature for 1 h. Then, the membranes were incubated with anti-ARID1A (1:500), anti-AKT (1:1,000) and anti-β-actin (1:5,000) primary antibodies at 4°C overnight, followed by incubation with a horseradish peroxide-conjugated mouse IgG secondary antibody (ab190475; 1:10,000; Abcam) or rabbit IgG secondary antibody (ab190495; 1:10,000; Abcam) at room temperature for 2 h. Proteins were visualized using an enhanced chemiluminescence kit (GE Healthcare Life Sciences, Piscataway, NJ, USA), and the protein band intensity was quantified via densitometric analysis using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

    Techniques: Expressing, Western Blot, Standard Deviation

    Kaempferol inactivates PI3K/AKT/mTOR signaling pathway in HepG2 cells by down-regulating miR-21. (a) Relative expressions of PTEN in HepG2 cells after 50 μM kaempferol treatment and/or miR-21 mimic transfection were assessed using quantitative reverse transcription PCR (qRT-PCR). (b) Western blotting was conducted to analyze the expressions of t-PI3K, p-PI3K, t-AKT, p-AKT, t-mTOR, p-mTOR, t-S6 K, and p-S6 K in HepG2 cells after 50 μM kaempferol treatment and/or miR-21 mimic transfection. miR-21: microRNA-21; PTEN: phosphatase and tensin homologue; NC: negative control; PI3K: phosphatidylinositol 3-kinase; AKT: protein kinase 3; mTOR: mechanistic target of rapamycin. * P

    Journal: International Journal of Immunopathology and Pharmacology

    Article Title: Kaempferol inhibits proliferation, migration, and invasion of liver cancer HepG2 cells by down-regulation of microRNA-21

    doi: 10.1177/2058738418814341

    Figure Lengend Snippet: Kaempferol inactivates PI3K/AKT/mTOR signaling pathway in HepG2 cells by down-regulating miR-21. (a) Relative expressions of PTEN in HepG2 cells after 50 μM kaempferol treatment and/or miR-21 mimic transfection were assessed using quantitative reverse transcription PCR (qRT-PCR). (b) Western blotting was conducted to analyze the expressions of t-PI3K, p-PI3K, t-AKT, p-AKT, t-mTOR, p-mTOR, t-S6 K, and p-S6 K in HepG2 cells after 50 μM kaempferol treatment and/or miR-21 mimic transfection. miR-21: microRNA-21; PTEN: phosphatase and tensin homologue; NC: negative control; PI3K: phosphatidylinositol 3-kinase; AKT: protein kinase 3; mTOR: mechanistic target of rapamycin. * P

    Article Snippet: Anti-Cyclin D1 antibody (ab21699), anti-pro-caspase 3 antibody (ab90437), anti-cleaved-caspase 3 antibody (ab2302), anti-pro-caspase 9 antibody (ab32068), anti-cleaved-caspase 9 antibody (ab2324), anti-Bcl-2 antibody (ab59348), anti-Bax antibody (ab182733), anti-matrix metalloproteinase 2 (MMP-2) antibody (ab37150), anti-MMP-9 antibody (ab73734), anti-Vimentin antibody (ab137321), anti-t-phosphatidylinositol 3-kinase (PI3K) antibody (ab28356), anti-p-PI3K antibody (ab182651), anti-t-protein kinase 3 (AKT) antibody (ab8805), anti-p-AKT antibody (ab38449), anti-t-mechanistic target of rapamycin (mTOR) antibody (ab32028), anti-p-mTOR antibody (ab63552), anti-t-S6 K antibody (ab9366), anti-p-S6 K antibody (ab131459), and anti-β-actin antibody (ab8226) were all purchased from Abcam Biotechnology (Cambridge, MA, USA).

    Techniques: Transfection, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Negative Control

    Time-dependent activation of Akt and eNOS paralleling PTEN phosphorylation by GTN. Representative Western blots showing PTEN phosphorylation (Ser 380), eNOS phosphorylation (Ser 1177 or 1179), and Akt phosphorylation (Ser 473) in (A) BAEC and (B) HMEC treated with vehicle or 500 nM GTN for the indicated amounts of time. Vehicle was added for 10 min in (A) and 15 min in (B). Results show rapid and sustained eNOS phosphorylation at the activation site Ser 1177, Akt phosphorylation at the activation site Ser 473, and PTEN phosphorylation at the inhibitory site Ser 380 by 500 nM GTN. Band intensities for phosphorylated eNOS in BAEC and for the experiments performed with HMEC were quantified using ImageJ and the values are reported as density units relative to control; * P

    Journal: Free radical biology & medicine

    Article Title: Nitroglycerin drives endothelial nitric oxide synthase activation via the phosphatidylinositol 3-kinase/protein kinase B pathway

    doi: 10.1016/j.freeradbiomed.2011.09.020

    Figure Lengend Snippet: Time-dependent activation of Akt and eNOS paralleling PTEN phosphorylation by GTN. Representative Western blots showing PTEN phosphorylation (Ser 380), eNOS phosphorylation (Ser 1177 or 1179), and Akt phosphorylation (Ser 473) in (A) BAEC and (B) HMEC treated with vehicle or 500 nM GTN for the indicated amounts of time. Vehicle was added for 10 min in (A) and 15 min in (B). Results show rapid and sustained eNOS phosphorylation at the activation site Ser 1177, Akt phosphorylation at the activation site Ser 473, and PTEN phosphorylation at the inhibitory site Ser 380 by 500 nM GTN. Band intensities for phosphorylated eNOS in BAEC and for the experiments performed with HMEC were quantified using ImageJ and the values are reported as density units relative to control; * P

    Article Snippet: Anti-phosphorylated eNOS (Ser 1177) was from BD Life Science and was used at 1:1000 dilution; phosphorylated PTEN (Ser 380) was from Cell Signaling Technology (Beverley, MA, USA) and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and phosphorylated Akt (Ser 473) antibodies were from Abcam (Cambridge, MA, USA) and were used at 1:1000 dilution.

    Techniques: Activation Assay, Western Blot