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    Cell Signaling Technology Inc phosphorylated akt ser473
    Phosphorylated Akt Ser473, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 32625 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phosphorylated+akt+ser473/Phospho-Akt+(Ser473)+Antibody/pm42834605-79-45-48
    Average 99 stars, based on 32625 article reviews
    phosphorylated akt ser473 - by Bioz Stars, 2026-10
    99/100 stars

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    Incubation:

    Article Title: Ivabradine Attenuates Experimental Hepatic Fibrosis by Modulating Inflammatory and Apoptotic Signaling Pathways
    Article Snippet: .. The blots were then incubated overnight at 4 °C with gentle agitation using the primary antibodies PI3K p85 (Cell Signaling, Danvers, MA, USA, #4292, 85 kDa, 1:1000), phosphorylated PI3K p85/p55 (Tyr467/Tyr199/Tyr464) (Invitrogen, Carlsbad, CA, USA, #PIPA5121306, 85 kDa, 1:1000), AKT (Cell Signaling, #9272, 60 kDa, 1:1000), phosphorylated AKT (Ser473) (Cell Signaling, #9271, 60 kDa, 1:1000), and mTOR (Cell Signaling, #2972, 289 kDa, 1:1000). ..

    Article Title: TAT-PBX1 Reverses Hyperglycemia Through β-Cell Regeneration and Functional Restoration in an STZ-Induced Diabetic Model
    Article Snippet: .. Membranes were then incubated overnight at 4 °C with primary antibodies: PBX1 (HA721303, dilution ratio 1:1000, Huabio, Hangzhou, China), Caspase 3 (AF6311, dilution ratio 1:1000, Affinity, Shanghai, China), Cleaved Caspase 3 (9661, dilution ratio 1:1000, CST, Danvers, MA, USA), GK (DF8549, dilution ratio 1:1000, Affinity, China), Pancreatic and Duodenal Homeobox 1 (PDX1, 20989-1-AP, dilution ratio 1:1000, Proteintech, Wuhan, China), p53-binding protein 1 (53BP1, ab36823, dilution ratio 1:1000, Abcam, Cambridge, UK), PARP1 (9532, dilution ratio 1:1000, CST, USA), γ-H2A.X (2577, dilution ratio 1:1000, CST, USA), Insulin Receptor Substrate 1 (IRS1, 17509-1-AP, dilution ratio 1:1000, Proteintech, China), Phosphoinositide 3-kinase regulatory subunit p85 (PI3K p85, T40115 , dilution ratio 1:1000, Abmart, Shanghai, China), Phosphorylated PI3K p85 (p-PI3K p85, T40116 , dilution ratio 1:1000, Abmart, China), AKT (9272, dilution ratio 1:1000, CST, USA), Phosphorylated AKT Ser473 (p-AKT S473, 9271, dilution ratio 1:1000, CST, USA), 3-Phosphoinositide-dependent protein kinase-1 (PDK1, F1103, dilution ratio 1:1000, Selleck, Houston, TX, USA), Phosphorylated PDK1 Ser241 (p-PDK1 S241, F0401, dilution ratio 1:1000, Selleck, Shanghai, China), GSK3β (F0142, dilution ratio 1:1000, Selleck, USA), Phosphorylated GSK3β Ser9 (p-GSK3β S9, F0299, dilution ratio 1:1000, Selleck, USA), β-Actin (PSH03-63, dilution ratio 1:1000, Huabio, China). .. The next day, membranes were washed three times with TBST and incubated with HRP-conjugated secondary antibodies at room temperature for 2 h. Protein bands were visualized using ECL substrate, and band intensity was semi-quantified using ImageJ software (version 1.8.0.172, National Institutes of Health, Bethesda, MD, USA).

    Article Title: Ivabradine Attenuates Experimental Hepatic Fibrosis by Modulating Inflammatory and Apoptotic Signaling Pathways.
    Article Snippet: .. The blots were then incubated overnight at 4 ◦C with gentle agitation using the primary antibodies PI3K p85 (Cell Signaling, Danvers, MA, USA, #4292, 85 kDa, 1:1000), phosphorylated PI3K p85/p55 (Tyr467/Tyr199/Tyr464) (Invitrogen, Carlsbad, CA, USA, #PIPA5121306, 85 kDa, 1:1000), AKT (Cell Signaling, #9272, 60 kDa, 1:1000), phosphorylated AKT (Ser473) (Cell Signaling, #9271, 60 kDa, 1:1000), and mTOR (Cell Signaling, #2972, 289 kDa, 1:1000). ..

    Article Title: TAT-PBX1 Reverses Hyperglycemia Through β-Cell Regeneration and Functional Restoration in an STZ-Induced Diabetic Model.
    Article Snippet: .. Membranes were then incubated overnight at 4 ◦C with primary antibodies: PBX1 (HA721303, dilution ratio 1:1000, Huabio, Hangzhou, China), Caspase 3 (AF6311, dilution ratio 1:1000, Affinity, Shanghai, China), Cleaved Caspase 3 (9661, dilution ratio 1:1000, CST, Danvers, MA, USA), GK (DF8549, dilution ratio 1:1000, Affinity, China), Pancreatic and Duodenal Homeobox 1 (PDX1, 20989-1-AP, dilution ratio 1:1000, Proteintech, Wuhan, China), p53-binding protein 1 (53BP1, ab36823, dilution ratio 1:1000, Abcam, Cambridge, UK), PARP1 (9532, dilution ratio 1:1000, CST, USA), γ-H2A.X (2577, dilution ratio 1:1000, CST, USA), Insulin Receptor Substrate 1 (IRS1, 17509-1-AP, dilution ratio 1:1000, Proteintech, China), Phosphoinositide 3-kinase regulatory subunit p85 (PI3K p85, T40115, dilution ratio 1:1000, Abmart, Shanghai, China), Phosphorylated PI3K p85 (p-PI3K p85, T40116, dilution ratio 1:1000, Abmart, China), AKT (9272, dilution ratio 1:1000, CST, USA), Phosphorylated AKT Ser473 (p-AKT S473, 9271, dilution ratio 1:1000, CST, USA), 3-Phosphoinositide-dependent protein kinase-1 (PDK1, F1103, dilution ratio 1:1000, Selleck, Houston, TX, USA), Phosphorylated PDK1 Ser241 (p-PDK1 S241, F0401, dilution ratio 1:1000, Selleck, Shanghai, China), GSK3β (F0142, dilution ratio 1:1000, Selleck, USA), Phosphorylated GSK3β Ser9 (p-GSK3β S9, F0299, dilution ratio 1:1000, Selleck, USA), β-Actin (PSH03-63, dilution ratio 1:1000, Huabio, China). .. The next day, membranes were washed three times with TBST and incubated with HRP-conjugated secondary antibodies at room temperature for 2 h. Protein bands https://doi.org/10.3390/ph19010085 were visualized using ECL substrate, and band intensity was semi-quantified using ImageJ software (version 1.8.0.172, National Institutes of Health, Bethesda, MD, USA).

    Article Title: Poly(ADP-ribose) Polymerase Inhibitor Talazoparib Inhibits DNA Damage Repair in Breast Cancer Cell Lines With or Without BRCA1/2 Mutation.
    Article Snippet: The cells were collected and fractionated using a Subcellular Protein Fraction Kit for Cultured Cells (Thermo Fisher Scientific Inc.) according to the manufacturer’s instructions. .. Separated proteins were transferred onto nitrocellulose membranes, and the blots were incubated overnight at 4°C with the following primary antibodies: PARP1 (sc-8007; Santa Cruz Biotechnology, Santa Cruz, USA), Lamin B1(sc-374015), Chk1 (sc-7898), Chk2 (sc-5278), Cyclin B1 (sc-245), TopBP1 (sc-271043), Rad9 (sc-8324), Hus1 (sc-30543), Rad51 (sc-377467), phosphorylated Akt (Ser473) (CST 9271; Cell Signaling Technology, Beverly, USA), Akt (CST 9272), phosphorylated Erk (p-Erk; CST 9101), Erk (CST 9102), caspase-3 (CST 9662), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; CST 2118), γ-H2AX (CST 9718), phosphorylated Chk1 (Ser345) (CST 2348), phosphorylated Chk2 (Thr68) (CST 2661), phosphorylated-ATR (Ser428) (CST 2853), ATR (CST 2790), MRE11 (CST 4847), NBS1 (CST 14956), histone H3 (ab1791; Abcam, Cambridge, UK), PAR (ab14459), RPA32 (ab2175), Rad50 (ab89), PARP (556494; BD Pharmingen, San Jose, USA), phosphorylated RPA (p-RPA; Ser4/Ser8) (A300-245A; Bethyl Laboratories, Montgomery, 4/15 Talazoparib Inhibits DNA Repair Regardless of BRCA1/2 Mutation Status https://doi.org/10.4048/jbc.2026.0085https://ejbc.kr Pr ov isi o al P ov isi on al USA), and Rad1 (NB100-56320; Novus Biologicals, Centennial, USA). ..

    Gentle:

    Article Title: Ivabradine Attenuates Experimental Hepatic Fibrosis by Modulating Inflammatory and Apoptotic Signaling Pathways
    Article Snippet: .. The blots were then incubated overnight at 4 °C with gentle agitation using the primary antibodies PI3K p85 (Cell Signaling, Danvers, MA, USA, #4292, 85 kDa, 1:1000), phosphorylated PI3K p85/p55 (Tyr467/Tyr199/Tyr464) (Invitrogen, Carlsbad, CA, USA, #PIPA5121306, 85 kDa, 1:1000), AKT (Cell Signaling, #9272, 60 kDa, 1:1000), phosphorylated AKT (Ser473) (Cell Signaling, #9271, 60 kDa, 1:1000), and mTOR (Cell Signaling, #2972, 289 kDa, 1:1000). ..

    Article Title: Ivabradine Attenuates Experimental Hepatic Fibrosis by Modulating Inflammatory and Apoptotic Signaling Pathways.
    Article Snippet: .. The blots were then incubated overnight at 4 ◦C with gentle agitation using the primary antibodies PI3K p85 (Cell Signaling, Danvers, MA, USA, #4292, 85 kDa, 1:1000), phosphorylated PI3K p85/p55 (Tyr467/Tyr199/Tyr464) (Invitrogen, Carlsbad, CA, USA, #PIPA5121306, 85 kDa, 1:1000), AKT (Cell Signaling, #9272, 60 kDa, 1:1000), phosphorylated AKT (Ser473) (Cell Signaling, #9271, 60 kDa, 1:1000), and mTOR (Cell Signaling, #2972, 289 kDa, 1:1000). ..

    Hemagglutination Assay:

    Article Title: Novel NOTCH2-NTRK1 fusion confers osimertinib resistance in EGFR-mutant non-small cell lung cancer by interacting with EGFR
    Article Snippet: BEAS-2B cells were grown in a DMEM medium, while H1650 and H1975 cells were cultured in an RPMI-1640 medium, plus with 10 %FBS, which were sourced from HyClone. .. Antibodies for total AKT (#4691), phosphorylated AKT (Ser473) (#4060), total ERK (#4695), phosphorylated ERK (Thr202/Tyr204) (#4370), HA (C29F4) (#3724), EGFR (D38B1) (#4267), phosphorylated EGFR (Thr669) (D2F1) (#8808), Met (D1C2) (#8198) and phosphorylated TRKA (Tyr490) (#4619) were purchased from Cell Signaling Technology. .. The anti-GAPDH antibody (#47724) was obtained from Santa Cruz Biotechnology.

    Article Title: Novel NOTCH2-NTRK1 fusion confers osimertinib resistance in EGFR-mutant non-small cell lung cancer by interacting with EGFR.
    Article Snippet: BEAS-2B cells were grown in a DMEM medium, while H1650 and H1975 cells were cultured in an RPMI-1640 medium, plus with 10 %FBS, which were sourced from HyClone. .. Antibodies for total AKT (#4691), phosphorylated AKT (Ser473) (#4060), total ERK (#4695), phosphorylated ERK (Thr202/Tyr204) (#4370), HA (C29F4) (#3724), EGFR (D38B1) (#4267), phosphorylated EGFR (Thr669) (D2F1) (#8808), Met (D1C2) (#8198) and phosphorylated TRKA (Tyr490) (#4619) were purchased from Cell Signaling Technology. .. The anti-GAPDH antibody (#47724) was obtained from Santa Cruz Biotechnology.

    Cell-Signaling:

    Article Title: Novel NOTCH2-NTRK1 fusion confers osimertinib resistance in EGFR-mutant non-small cell lung cancer by interacting with EGFR
    Article Snippet: BEAS-2B cells were grown in a DMEM medium, while H1650 and H1975 cells were cultured in an RPMI-1640 medium, plus with 10 %FBS, which were sourced from HyClone. .. Antibodies for total AKT (#4691), phosphorylated AKT (Ser473) (#4060), total ERK (#4695), phosphorylated ERK (Thr202/Tyr204) (#4370), HA (C29F4) (#3724), EGFR (D38B1) (#4267), phosphorylated EGFR (Thr669) (D2F1) (#8808), Met (D1C2) (#8198) and phosphorylated TRKA (Tyr490) (#4619) were purchased from Cell Signaling Technology. .. The anti-GAPDH antibody (#47724) was obtained from Santa Cruz Biotechnology.

    Article Title: Novel NOTCH2-NTRK1 fusion confers osimertinib resistance in EGFR-mutant non-small cell lung cancer by interacting with EGFR.
    Article Snippet: BEAS-2B cells were grown in a DMEM medium, while H1650 and H1975 cells were cultured in an RPMI-1640 medium, plus with 10 %FBS, which were sourced from HyClone. .. Antibodies for total AKT (#4691), phosphorylated AKT (Ser473) (#4060), total ERK (#4695), phosphorylated ERK (Thr202/Tyr204) (#4370), HA (C29F4) (#3724), EGFR (D38B1) (#4267), phosphorylated EGFR (Thr669) (D2F1) (#8808), Met (D1C2) (#8198) and phosphorylated TRKA (Tyr490) (#4619) were purchased from Cell Signaling Technology. .. The anti-GAPDH antibody (#47724) was obtained from Santa Cruz Biotechnology.

    Article Title: Poly(ADP-ribose) Polymerase Inhibitor Talazoparib Inhibits DNA Damage Repair in Breast Cancer Cell Lines With or Without BRCA1/2 Mutation.
    Article Snippet: The cells were collected and fractionated using a Subcellular Protein Fraction Kit for Cultured Cells (Thermo Fisher Scientific Inc.) according to the manufacturer’s instructions. .. Separated proteins were transferred onto nitrocellulose membranes, and the blots were incubated overnight at 4°C with the following primary antibodies: PARP1 (sc-8007; Santa Cruz Biotechnology, Santa Cruz, USA), Lamin B1(sc-374015), Chk1 (sc-7898), Chk2 (sc-5278), Cyclin B1 (sc-245), TopBP1 (sc-271043), Rad9 (sc-8324), Hus1 (sc-30543), Rad51 (sc-377467), phosphorylated Akt (Ser473) (CST 9271; Cell Signaling Technology, Beverly, USA), Akt (CST 9272), phosphorylated Erk (p-Erk; CST 9101), Erk (CST 9102), caspase-3 (CST 9662), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; CST 2118), γ-H2AX (CST 9718), phosphorylated Chk1 (Ser345) (CST 2348), phosphorylated Chk2 (Thr68) (CST 2661), phosphorylated-ATR (Ser428) (CST 2853), ATR (CST 2790), MRE11 (CST 4847), NBS1 (CST 14956), histone H3 (ab1791; Abcam, Cambridge, UK), PAR (ab14459), RPA32 (ab2175), Rad50 (ab89), PARP (556494; BD Pharmingen, San Jose, USA), phosphorylated RPA (p-RPA; Ser4/Ser8) (A300-245A; Bethyl Laboratories, Montgomery, 4/15 Talazoparib Inhibits DNA Repair Regardless of BRCA1/2 Mutation Status https://doi.org/10.4048/jbc.2026.0085https://ejbc.kr Pr ov isi o al P ov isi on al USA), and Rad1 (NB100-56320; Novus Biologicals, Centennial, USA). ..

    Recombinase Polymerase Amplification:

    Article Title: Poly(ADP-ribose) Polymerase Inhibitor Talazoparib Inhibits DNA Damage Repair in Breast Cancer Cell Lines With or Without BRCA1/2 Mutation.
    Article Snippet: The cells were collected and fractionated using a Subcellular Protein Fraction Kit for Cultured Cells (Thermo Fisher Scientific Inc.) according to the manufacturer’s instructions. .. Separated proteins were transferred onto nitrocellulose membranes, and the blots were incubated overnight at 4°C with the following primary antibodies: PARP1 (sc-8007; Santa Cruz Biotechnology, Santa Cruz, USA), Lamin B1(sc-374015), Chk1 (sc-7898), Chk2 (sc-5278), Cyclin B1 (sc-245), TopBP1 (sc-271043), Rad9 (sc-8324), Hus1 (sc-30543), Rad51 (sc-377467), phosphorylated Akt (Ser473) (CST 9271; Cell Signaling Technology, Beverly, USA), Akt (CST 9272), phosphorylated Erk (p-Erk; CST 9101), Erk (CST 9102), caspase-3 (CST 9662), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; CST 2118), γ-H2AX (CST 9718), phosphorylated Chk1 (Ser345) (CST 2348), phosphorylated Chk2 (Thr68) (CST 2661), phosphorylated-ATR (Ser428) (CST 2853), ATR (CST 2790), MRE11 (CST 4847), NBS1 (CST 14956), histone H3 (ab1791; Abcam, Cambridge, UK), PAR (ab14459), RPA32 (ab2175), Rad50 (ab89), PARP (556494; BD Pharmingen, San Jose, USA), phosphorylated RPA (p-RPA; Ser4/Ser8) (A300-245A; Bethyl Laboratories, Montgomery, 4/15 Talazoparib Inhibits DNA Repair Regardless of BRCA1/2 Mutation Status https://doi.org/10.4048/jbc.2026.0085https://ejbc.kr Pr ov isi o al P ov isi on al USA), and Rad1 (NB100-56320; Novus Biologicals, Centennial, USA). ..

    Mutagenesis:

    Article Title: Poly(ADP-ribose) Polymerase Inhibitor Talazoparib Inhibits DNA Damage Repair in Breast Cancer Cell Lines With or Without BRCA1/2 Mutation.
    Article Snippet: The cells were collected and fractionated using a Subcellular Protein Fraction Kit for Cultured Cells (Thermo Fisher Scientific Inc.) according to the manufacturer’s instructions. .. Separated proteins were transferred onto nitrocellulose membranes, and the blots were incubated overnight at 4°C with the following primary antibodies: PARP1 (sc-8007; Santa Cruz Biotechnology, Santa Cruz, USA), Lamin B1(sc-374015), Chk1 (sc-7898), Chk2 (sc-5278), Cyclin B1 (sc-245), TopBP1 (sc-271043), Rad9 (sc-8324), Hus1 (sc-30543), Rad51 (sc-377467), phosphorylated Akt (Ser473) (CST 9271; Cell Signaling Technology, Beverly, USA), Akt (CST 9272), phosphorylated Erk (p-Erk; CST 9101), Erk (CST 9102), caspase-3 (CST 9662), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; CST 2118), γ-H2AX (CST 9718), phosphorylated Chk1 (Ser345) (CST 2348), phosphorylated Chk2 (Thr68) (CST 2661), phosphorylated-ATR (Ser428) (CST 2853), ATR (CST 2790), MRE11 (CST 4847), NBS1 (CST 14956), histone H3 (ab1791; Abcam, Cambridge, UK), PAR (ab14459), RPA32 (ab2175), Rad50 (ab89), PARP (556494; BD Pharmingen, San Jose, USA), phosphorylated RPA (p-RPA; Ser4/Ser8) (A300-245A; Bethyl Laboratories, Montgomery, 4/15 Talazoparib Inhibits DNA Repair Regardless of BRCA1/2 Mutation Status https://doi.org/10.4048/jbc.2026.0085https://ejbc.kr Pr ov isi o al P ov isi on al USA), and Rad1 (NB100-56320; Novus Biologicals, Centennial, USA). ..



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    Image Search Results


    Local IL−10 administration induces Foxp3 expression and suppresses inflammatory signaling pathways in adipose SVFs of Lepr db/db mice. PBS or IL−10 (10, 50, or 100 ng) was injected directly into adipose tissue of Lepr db/db mice. SVFs were isolated 7 days after injection. ( A ) mRNA expression of inflammatory cytokines ( IL − 6 , IL-33 , CCL2 , IL − 1β , TNF − α ) and Foxp3 was quantified by Q-PCR. ( B ) Protein expression of phosphorylated and total JNK and NF-κB was evaluated by Western blotting to assess inflammatory signaling activity. Data are presented as mean ± SEM. n = 5 mice per group. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: International Journal of Molecular Sciences

    Article Title: MyD88 Inhibition Ameliorates Diabetes-Induced Hepatic Inflammation and Gluconeogenesis Through Adipose IL-10 Induction

    doi: 10.3390/ijms27062883

    Figure Lengend Snippet: Local IL−10 administration induces Foxp3 expression and suppresses inflammatory signaling pathways in adipose SVFs of Lepr db/db mice. PBS or IL−10 (10, 50, or 100 ng) was injected directly into adipose tissue of Lepr db/db mice. SVFs were isolated 7 days after injection. ( A ) mRNA expression of inflammatory cytokines ( IL − 6 , IL-33 , CCL2 , IL − 1β , TNF − α ) and Foxp3 was quantified by Q-PCR. ( B ) Protein expression of phosphorylated and total JNK and NF-κB was evaluated by Western blotting to assess inflammatory signaling activity. Data are presented as mean ± SEM. n = 5 mice per group. * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: Protein expression of phosphorylated Akt (pAkt; Cell Signaling, #4060), Akt (#4691), JNK (#9252), pJNK (#9251), ERK (#4695), and pERK (#9101) was determined by Western blotting.

    Techniques: Expressing, Protein-Protein interactions, Injection, Isolation, Western Blot, Activity Assay

    IL−10 administration suppresses hepatic ICAM , TNF − α , IL − 6 , DPP4 , and iNOS mRNA expression and activates insulin-related signaling pathways in Lepr db/db mice. PBS or IL-10 (10, 50, or 100 ng) was injected into adipose tissue of Lepr db/db mice, and livers were harvested 7 days later. ( A ) Hepatic mRNA expression of ICAM , FGF21 , IL − 1β , TNF − α , DPP4 , and iNOS was determined by Q-PCR. ( B ) Protein expression of phosphorylated and total Akt, STAT3, and ERK was assessed by Western blotting. ( C ) Quantification of the pAkt/Akt and pSTAT3/STAT3 ratio. Data are presented as mean ± SEM. n = 4 mice per group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Journal: International Journal of Molecular Sciences

    Article Title: MyD88 Inhibition Ameliorates Diabetes-Induced Hepatic Inflammation and Gluconeogenesis Through Adipose IL-10 Induction

    doi: 10.3390/ijms27062883

    Figure Lengend Snippet: IL−10 administration suppresses hepatic ICAM , TNF − α , IL − 6 , DPP4 , and iNOS mRNA expression and activates insulin-related signaling pathways in Lepr db/db mice. PBS or IL-10 (10, 50, or 100 ng) was injected into adipose tissue of Lepr db/db mice, and livers were harvested 7 days later. ( A ) Hepatic mRNA expression of ICAM , FGF21 , IL − 1β , TNF − α , DPP4 , and iNOS was determined by Q-PCR. ( B ) Protein expression of phosphorylated and total Akt, STAT3, and ERK was assessed by Western blotting. ( C ) Quantification of the pAkt/Akt and pSTAT3/STAT3 ratio. Data are presented as mean ± SEM. n = 4 mice per group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

    Article Snippet: Protein expression of phosphorylated Akt (pAkt; Cell Signaling, #4060), Akt (#4691), JNK (#9252), pJNK (#9251), ERK (#4695), and pERK (#9101) was determined by Western blotting.

    Techniques: Expressing, Protein-Protein interactions, Injection, Western Blot

    IL−10 administration reduces hepatic gluconeogenic gene expression, enhances Akt activation, and improves glucose tolerance in Lepr db/db mice. PBS or IL-10 (10 or 50 ng) was injected into inguinal white adipose tissue (WAT) of Lepr db/db mice, the liver was harvested 7 days after injection and subjected to Q-PCR analysis to determine the expression of G6pc and Pck1 mRNA ( A ). ( B ) One week after injection, mice were treated with insulin (1.25 mU/g body weight) for 20 min, followed by isolation of SVFs and Western blot analysis of phosphorylated and total Akt. ( C ) Quantification of the pAkt/Akt ratio. ( D ) Glucose tolerance tests were performed by intraperitoneal glucose administration (1 g/kg body weight), with blood glucose measured at baseline and at 15 min intervals for 2 h. Data are presented as mean ± SEM. n = 4 mice per group. * p < 0.05, ** p < 0.01; # p < 0.05 compared with PBS-treated Lepr db/db mice.

    Journal: International Journal of Molecular Sciences

    Article Title: MyD88 Inhibition Ameliorates Diabetes-Induced Hepatic Inflammation and Gluconeogenesis Through Adipose IL-10 Induction

    doi: 10.3390/ijms27062883

    Figure Lengend Snippet: IL−10 administration reduces hepatic gluconeogenic gene expression, enhances Akt activation, and improves glucose tolerance in Lepr db/db mice. PBS or IL-10 (10 or 50 ng) was injected into inguinal white adipose tissue (WAT) of Lepr db/db mice, the liver was harvested 7 days after injection and subjected to Q-PCR analysis to determine the expression of G6pc and Pck1 mRNA ( A ). ( B ) One week after injection, mice were treated with insulin (1.25 mU/g body weight) for 20 min, followed by isolation of SVFs and Western blot analysis of phosphorylated and total Akt. ( C ) Quantification of the pAkt/Akt ratio. ( D ) Glucose tolerance tests were performed by intraperitoneal glucose administration (1 g/kg body weight), with blood glucose measured at baseline and at 15 min intervals for 2 h. Data are presented as mean ± SEM. n = 4 mice per group. * p < 0.05, ** p < 0.01; # p < 0.05 compared with PBS-treated Lepr db/db mice.

    Article Snippet: Protein expression of phosphorylated Akt (pAkt; Cell Signaling, #4060), Akt (#4691), JNK (#9252), pJNK (#9251), ERK (#4695), and pERK (#9101) was determined by Western blotting.

    Techniques: Gene Expression, Activation Assay, Injection, Expressing, Isolation, Western Blot

    KSM inhibits CHI3L1-stimulated Erk and Akt activation and the antiapoptotic effect of CHI3L1 in macrophages. ( A ) AMJ-C11 mouse lung macrophages were stimulated with recombinant (r) CHI3L1 overnight, and the activation of Erk and Akt was detected by Western blot evaluations. pErk, phosphorylated Erk; T-Erk, total Erk; pAkt, phosphorylated Akt, T-Akt, total Akt. Right panel, densitometric quantitation on the blots of pErk and pAkt. ( B ) After exposure of AMJ-C11 cells with H 2 O 2 (1 mM, 24 hours) with and without rCHI3L1 and KSM treatment, cellular apoptosis (% of Annexin V+ cells) was measured by FACS evaluations. ( A and B ) are representatives of 3 independent experiments. Right panel, the values of Annexin 5 (+) cells (%) are the mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001 (One-Way ANOVA, multiple comparisons).

    Journal: ImmunoTargets and Therapy

    Article Title: Kasugamycin Inhibits Melanoma Lung Metastasis and CHI3L1-Driven M2-Like Tumor-Associated Macrophage Differentiation

    doi: 10.2147/ITT.S563951

    Figure Lengend Snippet: KSM inhibits CHI3L1-stimulated Erk and Akt activation and the antiapoptotic effect of CHI3L1 in macrophages. ( A ) AMJ-C11 mouse lung macrophages were stimulated with recombinant (r) CHI3L1 overnight, and the activation of Erk and Akt was detected by Western blot evaluations. pErk, phosphorylated Erk; T-Erk, total Erk; pAkt, phosphorylated Akt, T-Akt, total Akt. Right panel, densitometric quantitation on the blots of pErk and pAkt. ( B ) After exposure of AMJ-C11 cells with H 2 O 2 (1 mM, 24 hours) with and without rCHI3L1 and KSM treatment, cellular apoptosis (% of Annexin V+ cells) was measured by FACS evaluations. ( A and B ) are representatives of 3 independent experiments. Right panel, the values of Annexin 5 (+) cells (%) are the mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001 (One-Way ANOVA, multiple comparisons).

    Article Snippet: Protein lysates (25 μg) from lung tissues or cells were subjected to SDS-PAGE, transferred to membranes, and immunoblotted with primary antibodies against CD163 (#PA5-109327, Thermo Fisher Scientific), CD206 (E6T5J, #24595S, Cell Signaling Technology), phosphorylated AKT (p-Akt) (193H12, #4058S, Cell Signaling Technology), and total Akt (11E7, #4685S, Cell Signaling Technology), phosphorylated EGFR (Tyr1068) (p-EGFR) (#44-788G, Thermo Fisher Scientific), EGFR (D38B1, #4267S, Cell Signaling Technology), phosphorylated Erk (p-Erk) (#9101S, Cell Signaling Technology), total Erk (#9102S, Cell Signaling Technology), phosphorylated STAT3 (Tyr705) (p-STAT3) (D3A7, #9145, Cell Signaling Technology), total STAT3 (79D7, #4904, Cell Signaling Technology) and b-actin (C4, #sc-47778 HRP, Santa Cruz Biotechnology).

    Techniques: Activation Assay, Recombinant, Western Blot, Quantitation Assay