anti acc2  (Cell Signaling Technology Inc)

Journal: Experimental and Therapeutic Medicine

Article Title: Lithium downregulates phosphorylated acetyl‑CoA carboxylase 2 and attenuates mitochondrial fatty acid utilization and oxidative stress in cardiomyocytes

doi: 10.3892/etm.2024.12413

Figure Lengend Snippet: Expression ACC2 and GSK-3β in H9c2 cardiomyocytes treated with LiCl. (A) Compared with control cells (n=5), H9c2 cells treated with LiCl at 0.3 mM (n=5) for 48 h exhibited downregulated expression of p-ACC2. However, LiCl at 0.1 (n=5) or 1.0 mM (n=5) had no significant effect on the expression of p-ACC2. Additionally, the expression of total ACC2 did not significantly differ between the control cells and those treated with LiCl at concentrations of 0.1, 0.3 and 1 mM. (B) H9c2 cells treated with LiCl at 0.3 mM (n=5) for 48 h exhibited upregulated expression of p-GSK-3β relative to the control cells (n=5). However, LiCl at 0.1 (n=5) and 1.0 mM (n=5) did not significantly affect the expression of p-GSK-3β. The expression of total GSK-3β did not significantly differ between the control cells and those treated with LiCl at concentrations of 0.1, 0.3 and 1 mM. * P<0.05. ACC2, acetyl-CoA carboxylase 2; GSK-3β, glycogen synthase kinase-3 beta; p-, phosphorylated; CON, control.

Article Snippet: Following this blocking procedure, the membranes were incubated overnight at 4˚C with specific antibodies against total ACC2 (1:2,000; monoclonal; cat. no. ab45174; Abcam), p-(p-)ACC2 (1:500; polyclonal; cat. no. 07303; Millipore), total AMP-activated protein kinase (AMPK; 1:500; monoclonal; cat. no. 5831; Cell Signaling), p-AMPK (1:1,000; polyclonal; cat. no. 07681; Millipore), calcineurin (1:10,000; monoclonal; cat. no. ab109412; Abcam), total GSK-3β (1:1,000; monoclonal; cat. no. 9315; Cell Signaling), p-GSK-3β (1:1,000; polyclonal; cat. no. 9336; Cell Signaling) and GLUT4 (1:500; monoclonal; cat. no. sc-53566; Santa Cruz).

Techniques: Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Lithium downregulates phosphorylated acetyl‑CoA carboxylase 2 and attenuates mitochondrial fatty acid utilization and oxidative stress in cardiomyocytes

doi: 10.3892/etm.2024.12413

Figure Lengend Snippet: Expression of ACC2 in H9c2 cardiomyocytes treated with a glycogen synthase kinase-3 beta GSK-3β inhibitor. Cotreatment with TWS119 (8 µM) and LiCl (0.3 mM) downregulated the expression of p-ACC2 in H9c2 cells to a similar extent as did treatment with TWS119 (8 µM) alone. The expression of total ACC2 did not significantly differ between the control cells and those treated with TWS119 (8 µM alone or cotreatment with TWS119 [8 µM] and LiCl [0.3 mM]). *** P<0.001. ACC2, acetyl-CoA carboxylase 2; GSK-3β, glycogen synthase kinase-3 beta; p-, phosphorylated; CON, control.

Article Snippet: Following this blocking procedure, the membranes were incubated overnight at 4˚C with specific antibodies against total ACC2 (1:2,000; monoclonal; cat. no. ab45174; Abcam), p-(p-)ACC2 (1:500; polyclonal; cat. no. 07303; Millipore), total AMP-activated protein kinase (AMPK; 1:500; monoclonal; cat. no. 5831; Cell Signaling), p-AMPK (1:1,000; polyclonal; cat. no. 07681; Millipore), calcineurin (1:10,000; monoclonal; cat. no. ab109412; Abcam), total GSK-3β (1:1,000; monoclonal; cat. no. 9315; Cell Signaling), p-GSK-3β (1:1,000; polyclonal; cat. no. 9336; Cell Signaling) and GLUT4 (1:500; monoclonal; cat. no. sc-53566; Santa Cruz).

Techniques: Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Lithium downregulates phosphorylated acetyl‑CoA carboxylase 2 and attenuates mitochondrial fatty acid utilization and oxidative stress in cardiomyocytes

doi: 10.3892/etm.2024.12413

Figure Lengend Snippet: Illustration of the proposed mechanisms underlying lithium's effects on mitochondrial fatty acid oxidation and oxidative stress in cardiomyocytes. Low-dose lithium downregulates GSK-3β activity in cardiomyocytes and activates ACC2, thereby upregulating malonyl-CoA expression, which in turn leads to the inhibition on CPT1 activity, transportation of fatty acyl-CoAs into mitochondria for β-oxidation and generation of ROS and proton leakage. GLUT4, glucose transporter type 4; ACC2, acetyl-CoA carboxylase 2; CPT1, carnitine palmitoyltransferase 1; ROS, reactive oxygen species.

Article Snippet: Following this blocking procedure, the membranes were incubated overnight at 4˚C with specific antibodies against total ACC2 (1:2,000; monoclonal; cat. no. ab45174; Abcam), p-(p-)ACC2 (1:500; polyclonal; cat. no. 07303; Millipore), total AMP-activated protein kinase (AMPK; 1:500; monoclonal; cat. no. 5831; Cell Signaling), p-AMPK (1:1,000; polyclonal; cat. no. 07681; Millipore), calcineurin (1:10,000; monoclonal; cat. no. ab109412; Abcam), total GSK-3β (1:1,000; monoclonal; cat. no. 9315; Cell Signaling), p-GSK-3β (1:1,000; polyclonal; cat. no. 9336; Cell Signaling) and GLUT4 (1:500; monoclonal; cat. no. sc-53566; Santa Cruz).

Techniques: Activity Assay, Expressing, Inhibition

Journal: Experimental and Therapeutic Medicine

Article Title: Lithium downregulates phosphorylated acetyl‑CoA carboxylase 2 and attenuates mitochondrial fatty acid utilization and oxidative stress in cardiomyocytes

doi: 10.3892/etm.2024.12413

Figure Lengend Snippet: Expression ACC2 and GSK-3β in H9c2 cardiomyocytes treated with LiCl. (A) Compared with control cells (n=5), H9c2 cells treated with LiCl at 0.3 mM (n=5) for 48 h exhibited downregulated expression of p-ACC2. However, LiCl at 0.1 (n=5) or 1.0 mM (n=5) had no significant effect on the expression of p-ACC2. Additionally, the expression of total ACC2 did not significantly differ between the control cells and those treated with LiCl at concentrations of 0.1, 0.3 and 1 mM. (B) H9c2 cells treated with LiCl at 0.3 mM (n=5) for 48 h exhibited upregulated expression of p-GSK-3β relative to the control cells (n=5). However, LiCl at 0.1 (n=5) and 1.0 mM (n=5) did not significantly affect the expression of p-GSK-3β. The expression of total GSK-3β did not significantly differ between the control cells and those treated with LiCl at concentrations of 0.1, 0.3 and 1 mM. * P<0.05. ACC2, acetyl-CoA carboxylase 2; GSK-3β, glycogen synthase kinase-3 beta; p-, phosphorylated; CON, control.

Article Snippet: Following this blocking procedure, the membranes were incubated overnight at 4˚C with specific antibodies against total ACC2 (1:2,000; monoclonal; cat. no. ab45174; Abcam), p-(p-)ACC2 (1:500; polyclonal; cat. no. 07303; Millipore), total AMP-activated protein kinase (AMPK; 1:500; monoclonal; cat. no. 5831; Cell Signaling), p-AMPK (1:1,000; polyclonal; cat. no. 07681; Millipore), calcineurin (1:10,000; monoclonal; cat. no. ab109412; Abcam), total GSK-3β (1:1,000; monoclonal; cat. no. 9315; Cell Signaling), p-GSK-3β (1:1,000; polyclonal; cat. no. 9336; Cell Signaling) and GLUT4 (1:500; monoclonal; cat. no. sc-53566; Santa Cruz).

Techniques: Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Lithium downregulates phosphorylated acetyl‑CoA carboxylase 2 and attenuates mitochondrial fatty acid utilization and oxidative stress in cardiomyocytes

doi: 10.3892/etm.2024.12413

Figure Lengend Snippet: Expression of ACC2 in H9c2 cardiomyocytes treated with a glycogen synthase kinase-3 beta GSK-3β inhibitor. Cotreatment with TWS119 (8 µM) and LiCl (0.3 mM) downregulated the expression of p-ACC2 in H9c2 cells to a similar extent as did treatment with TWS119 (8 µM) alone. The expression of total ACC2 did not significantly differ between the control cells and those treated with TWS119 (8 µM alone or cotreatment with TWS119 [8 µM] and LiCl [0.3 mM]). *** P<0.001. ACC2, acetyl-CoA carboxylase 2; GSK-3β, glycogen synthase kinase-3 beta; p-, phosphorylated; CON, control.

Article Snippet: Following this blocking procedure, the membranes were incubated overnight at 4˚C with specific antibodies against total ACC2 (1:2,000; monoclonal; cat. no. ab45174; Abcam), p-(p-)ACC2 (1:500; polyclonal; cat. no. 07303; Millipore), total AMP-activated protein kinase (AMPK; 1:500; monoclonal; cat. no. 5831; Cell Signaling), p-AMPK (1:1,000; polyclonal; cat. no. 07681; Millipore), calcineurin (1:10,000; monoclonal; cat. no. ab109412; Abcam), total GSK-3β (1:1,000; monoclonal; cat. no. 9315; Cell Signaling), p-GSK-3β (1:1,000; polyclonal; cat. no. 9336; Cell Signaling) and GLUT4 (1:500; monoclonal; cat. no. sc-53566; Santa Cruz).

Techniques: Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Lithium downregulates phosphorylated acetyl‑CoA carboxylase 2 and attenuates mitochondrial fatty acid utilization and oxidative stress in cardiomyocytes

doi: 10.3892/etm.2024.12413

Figure Lengend Snippet: Illustration of the proposed mechanisms underlying lithium's effects on mitochondrial fatty acid oxidation and oxidative stress in cardiomyocytes. Low-dose lithium downregulates GSK-3β activity in cardiomyocytes and activates ACC2, thereby upregulating malonyl-CoA expression, which in turn leads to the inhibition on CPT1 activity, transportation of fatty acyl-CoAs into mitochondria for β-oxidation and generation of ROS and proton leakage. GLUT4, glucose transporter type 4; ACC2, acetyl-CoA carboxylase 2; CPT1, carnitine palmitoyltransferase 1; ROS, reactive oxygen species.

Article Snippet: Following this blocking procedure, the membranes were incubated overnight at 4˚C with specific antibodies against total ACC2 (1:2,000; monoclonal; cat. no. ab45174; Abcam), p-(p-)ACC2 (1:500; polyclonal; cat. no. 07303; Millipore), total AMP-activated protein kinase (AMPK; 1:500; monoclonal; cat. no. 5831; Cell Signaling), p-AMPK (1:1,000; polyclonal; cat. no. 07681; Millipore), calcineurin (1:10,000; monoclonal; cat. no. ab109412; Abcam), total GSK-3β (1:1,000; monoclonal; cat. no. 9315; Cell Signaling), p-GSK-3β (1:1,000; polyclonal; cat. no. 9336; Cell Signaling) and GLUT4 (1:500; monoclonal; cat. no. sc-53566; Santa Cruz).

Techniques: Activity Assay, Expressing, Inhibition

Journal: Heliyon

Article Title: miR-122/PPARβ axis is involved in hypoxic exercise and modulates fatty acid metabolism in skeletal muscle of obese rats

doi: 10.1016/j.heliyon.2024.e26572

Figure Lengend Snippet: Primers used for RT-qPCR analysis.

Article Snippet: The protein samples were then transferred to a PVDF membrane (0.45-μm pore size) at a constant pressure of 100 V for 10 min, followed by blocked overnight in Tris-buffered saline with 0.1% Tween-20 (TBST), which contained 5% bovine serum albumin, and then probed overnight at 4 °C with primary antibodies against PPARβ (1:2000; GTX113250) and FAS (1:1000; GTX13550) (both from GeneTex, Irvine, CA, USA) and ACC2 (1:500; A03668-2) and CPT1b (1:500; PB9491) (both from Boster Bio, Pleasanton, CA, USA).

Techniques: Amplification

Journal: Heliyon

Article Title: miR-122/PPARβ axis is involved in hypoxic exercise and modulates fatty acid metabolism in skeletal muscle of obese rats

doi: 10.1016/j.heliyon.2024.e26572

Figure Lengend Snippet: Expression of the lipid metabolism regulator PPARβ and downstream effectors (CPT1b, FAS, and ACC2). mRNA levels in obese rats under hypoxic conditions, with hypoxic training, and with regulated miR-122 expression were determined using qRT-PCR. OE, obese rats with miR-122 overexpression and hypoxic training; IE, obese rats with miR-122 depletion and hypoxic training; CE, obese rats with hypoxic training only; H, obese sedentary rats without regulation of miR-122 expression. Data are presented as mean ± SD. *p < 0.05, **p < 0.01 vs rats in group H; # p < 0.05, ## p < 0.01 vs rats in group CE; & p < 0.05, && p < 0.01 vs rats in group OE (1-way analysis of variance).

Article Snippet: The protein samples were then transferred to a PVDF membrane (0.45-μm pore size) at a constant pressure of 100 V for 10 min, followed by blocked overnight in Tris-buffered saline with 0.1% Tween-20 (TBST), which contained 5% bovine serum albumin, and then probed overnight at 4 °C with primary antibodies against PPARβ (1:2000; GTX113250) and FAS (1:1000; GTX13550) (both from GeneTex, Irvine, CA, USA) and ACC2 (1:500; A03668-2) and CPT1b (1:500; PB9491) (both from Boster Bio, Pleasanton, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Over Expression

Journal: Heliyon

Article Title: miR-122/PPARβ axis is involved in hypoxic exercise and modulates fatty acid metabolism in skeletal muscle of obese rats

doi: 10.1016/j.heliyon.2024.e26572

Figure Lengend Snippet: Expression of the lipid metabolism regulator PPARβ and downstream effectors (CPT1b, FAS, and ACC2). Protein levels in obese rats under hypoxic conditions, with hypoxic training, and with regulated miR-122 expression were determined using Western blot analysis. OE, obese rats with miR-122 overexpression and hypoxic training; IE, obese rats with miR-122 depletion and hypoxic training; CE, obese rats with hypoxic training only; H, obese sedentary rats without regulation of miR-122 expression. Data are presented as mean ± SD. *p < 0.05, **p < 0.01 vs rats in group H; # p < 0.05, ## p < 0.01 vs rats in group CE; & p < 0.05, && p < 0.01 vs rats in group OE (1-way analysis of variance).

Article Snippet: The protein samples were then transferred to a PVDF membrane (0.45-μm pore size) at a constant pressure of 100 V for 10 min, followed by blocked overnight in Tris-buffered saline with 0.1% Tween-20 (TBST), which contained 5% bovine serum albumin, and then probed overnight at 4 °C with primary antibodies against PPARβ (1:2000; GTX113250) and FAS (1:1000; GTX13550) (both from GeneTex, Irvine, CA, USA) and ACC2 (1:500; A03668-2) and CPT1b (1:500; PB9491) (both from Boster Bio, Pleasanton, CA, USA).

Techniques: Expressing, Western Blot, Over Expression

Journal: Heliyon

Article Title: miR-122/PPARβ axis is involved in hypoxic exercise and modulates fatty acid metabolism in skeletal muscle of obese rats

doi: 10.1016/j.heliyon.2024.e26572

Figure Lengend Snippet: Primers used for RT-qPCR analysis.

Article Snippet: The protein samples were then transferred to a PVDF membrane (0.45-μm pore size) at a constant pressure of 100 V for 10 min, followed by blocked overnight in Tris-buffered saline with 0.1% Tween-20 (TBST), which contained 5% bovine serum albumin, and then probed overnight at 4 °C with primary antibodies against PPARβ (1:2000; GTX113250) and FAS (1:1000; GTX13550) (both from GeneTex, Irvine, CA, USA) and ACC2 (1:500; A03668-2) and CPT1b (1:500; PB9491) (both from Boster Bio, Pleasanton, CA, USA).

Techniques: Amplification

Journal: Heliyon

Article Title: miR-122/PPARβ axis is involved in hypoxic exercise and modulates fatty acid metabolism in skeletal muscle of obese rats

doi: 10.1016/j.heliyon.2024.e26572

Figure Lengend Snippet: Expression of the lipid metabolism regulator PPARβ and downstream effectors (CPT1b, FAS, and ACC2). mRNA levels in obese rats under hypoxic conditions, with hypoxic training, and with regulated miR-122 expression were determined using qRT-PCR. OE, obese rats with miR-122 overexpression and hypoxic training; IE, obese rats with miR-122 depletion and hypoxic training; CE, obese rats with hypoxic training only; H, obese sedentary rats without regulation of miR-122 expression. Data are presented as mean ± SD. *p < 0.05, **p < 0.01 vs rats in group H; # p < 0.05, ## p < 0.01 vs rats in group CE; & p < 0.05, && p < 0.01 vs rats in group OE (1-way analysis of variance).

Article Snippet: The protein samples were then transferred to a PVDF membrane (0.45-μm pore size) at a constant pressure of 100 V for 10 min, followed by blocked overnight in Tris-buffered saline with 0.1% Tween-20 (TBST), which contained 5% bovine serum albumin, and then probed overnight at 4 °C with primary antibodies against PPARβ (1:2000; GTX113250) and FAS (1:1000; GTX13550) (both from GeneTex, Irvine, CA, USA) and ACC2 (1:500; A03668-2) and CPT1b (1:500; PB9491) (both from Boster Bio, Pleasanton, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Over Expression

Journal: Heliyon

Article Title: miR-122/PPARβ axis is involved in hypoxic exercise and modulates fatty acid metabolism in skeletal muscle of obese rats

doi: 10.1016/j.heliyon.2024.e26572

Figure Lengend Snippet: Expression of the lipid metabolism regulator PPARβ and downstream effectors (CPT1b, FAS, and ACC2). Protein levels in obese rats under hypoxic conditions, with hypoxic training, and with regulated miR-122 expression were determined using Western blot analysis. OE, obese rats with miR-122 overexpression and hypoxic training; IE, obese rats with miR-122 depletion and hypoxic training; CE, obese rats with hypoxic training only; H, obese sedentary rats without regulation of miR-122 expression. Data are presented as mean ± SD. *p < 0.05, **p < 0.01 vs rats in group H; # p < 0.05, ## p < 0.01 vs rats in group CE; & p < 0.05, && p < 0.01 vs rats in group OE (1-way analysis of variance).

Article Snippet: The protein samples were then transferred to a PVDF membrane (0.45-μm pore size) at a constant pressure of 100 V for 10 min, followed by blocked overnight in Tris-buffered saline with 0.1% Tween-20 (TBST), which contained 5% bovine serum albumin, and then probed overnight at 4 °C with primary antibodies against PPARβ (1:2000; GTX113250) and FAS (1:1000; GTX13550) (both from GeneTex, Irvine, CA, USA) and ACC2 (1:500; A03668-2) and CPT1b (1:500; PB9491) (both from Boster Bio, Pleasanton, CA, USA).

Techniques: Expressing, Western Blot, Over Expression