Membrane:Article Title: Tebuconazole Fungicide Induces Lipid Accumulation and Oxidative Stress in HepG2 Cells
Article Snippet: .. The membrane was incubated with the primary antibodies: anti-PPARα (1:1000 dilution; Santa Cruz, CA, USA), anti-PPARγ (1:3000 dilution; Cell Signaling Technology, Beverly, MA, USA), anti-cluster of differentiation 36 (CD36) (1:3000 dilution; Cell Signaling Technology, Beverly, MA, USA), anti-fatty acid transporter protein (FATP) 2 (1:3000 dilution; Abcam, Cambridge, MA, USA), anti-microsomal triglyceride transfer protein (MTTP) (1:3000 dilution; Abcam, Cambridge, MA, USA), anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:20,000 dilution; Merck Millipore, Darmstadt, Germany), and anti-Lamin B (1:5,000 dilution; Santa Cruz, CA, USA) at 4 °C from overnight to 3 days. .. After washing thrice with Tris-buffered saline with Tween 20, secondary antibodies (i.e., goat anti-rabbit IgG (1:5000 dilution; Enzo Life Sciences, Lausen, Switzerland) and donkey anti-goat IgG (1:5000 dilution; Santa Cruz, CA, USA) conjugated with horseradish peroxidase) were incubated for 1.5 h at 25 °C.
Incubation:Article Title: Tebuconazole Fungicide Induces Lipid Accumulation and Oxidative Stress in HepG2 Cells
Article Snippet: .. The membrane was incubated with the primary antibodies: anti-PPARα (1:1000 dilution; Santa Cruz, CA, USA), anti-PPARγ (1:3000 dilution; Cell Signaling Technology, Beverly, MA, USA), anti-cluster of differentiation 36 (CD36) (1:3000 dilution; Cell Signaling Technology, Beverly, MA, USA), anti-fatty acid transporter protein (FATP) 2 (1:3000 dilution; Abcam, Cambridge, MA, USA), anti-microsomal triglyceride transfer protein (MTTP) (1:3000 dilution; Abcam, Cambridge, MA, USA), anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:20,000 dilution; Merck Millipore, Darmstadt, Germany), and anti-Lamin B (1:5,000 dilution; Santa Cruz, CA, USA) at 4 °C from overnight to 3 days. .. After washing thrice with Tris-buffered saline with Tween 20, secondary antibodies (i.e., goat anti-rabbit IgG (1:5000 dilution; Enzo Life Sciences, Lausen, Switzerland) and donkey anti-goat IgG (1:5000 dilution; Santa Cruz, CA, USA) conjugated with horseradish peroxidase) were incubated for 1.5 h at 25 °C.
Staining:Article Title: AMPK-Dependent Inhibitory Phosphorylation of ACC Is Not Essential for Maintaining Myocardial Fatty Acid Oxidation
Article Snippet: .. Blotted proteins were reversibly visualized using MemCode stain (Pierce) and identified using the primary antibodies: anti-ACC (EMD Millipore, Billerica, MA), anti-AMPK2, anti-fatty acid transport protein 1 (FATP1), anti-peroxisome proliferator-activated receptor alpha (PPARα), anti-actin (Santa Cruz Biotechnology, Dallas, TX), anti-cluster of differentiation 36 (CD36) (generated in-house), anti-phospho ACC Ser79, anti-phospho AMPK Thr172, anti-adipose triglyceride lipase (ATGL), anti-phospho Akt Ser473, anti-phospho Akt Thr308, anti-Akt, ant-phospho p38 Thr108, anti-p38, anti-phospho protein kinase A (PKA) Thr197, anti-PKA (Cell Signaling Technology, Danvers, MA) antibodies. .. Immunoblots were developed using the Western Lightning Plus-ECL enhanced chemiluminescence substrate (Perkin Elmer, Waltham, MA).
Generated:Article Title: AMPK-Dependent Inhibitory Phosphorylation of ACC Is Not Essential for Maintaining Myocardial Fatty Acid Oxidation
Article Snippet: .. Blotted proteins were reversibly visualized using MemCode stain (Pierce) and identified using the primary antibodies: anti-ACC (EMD Millipore, Billerica, MA), anti-AMPK2, anti-fatty acid transport protein 1 (FATP1), anti-peroxisome proliferator-activated receptor alpha (PPARα), anti-actin (Santa Cruz Biotechnology, Dallas, TX), anti-cluster of differentiation 36 (CD36) (generated in-house), anti-phospho ACC Ser79, anti-phospho AMPK Thr172, anti-adipose triglyceride lipase (ATGL), anti-phospho Akt Ser473, anti-phospho Akt Thr308, anti-Akt, ant-phospho p38 Thr108, anti-p38, anti-phospho protein kinase A (PKA) Thr197, anti-PKA (Cell Signaling Technology, Danvers, MA) antibodies. .. Immunoblots were developed using the Western Lightning Plus-ECL enhanced chemiluminescence substrate (Perkin Elmer, Waltham, MA).
Western Blot:Article Title: Tofogliflozin attenuates liver steatosis and fibrosis in non-diabetic non-alcoholic steatohepatitis mice
Article Snippet: .. The primary antibodies used in immunoblotting included antibodies against acetyl-CoA carboxylase (ACC) (#3676, Cell Signaling Technology, MA, USA), cluster of differentiation 36 (CD36) (#28109, Cell Signaling Technology), carnitine palmitoyltransferase 1a (CPT1a) (ab234111, Abcam), fatty acid synthase (FASN) (#3180, Cell Signaling Technology), fatty acid transport protein 2 (FATP2) (#78771, Cell Signaling Technology), farnesoid X receptor (FXR) (#72105, Cell Signaling Technology), and sterol regulatory element binding protein-1c (SREBP-1c) (sc-13551, Santa Cruz Biotechnology, TX, USA). .. The membranes were washed with TBST and incubated with goat anti-rabbit (#31460, Thermo Fisher Scientific, MA, USA) or goat anti-mouse secondary antibodies (#31430, Thermo Fisher Scientific) at room temperature for 1 h. The blots were incubated with β-actin (#4970, Cell Signaling Technology) as a loading control.
Article Title: Tofogliflozin attenuates liver steatosis and fibrosis in non-diabetic non-alcoholic steatohepatitis mice.
Article Snippet: .. The primary antibodies used in immunoblotting included antibodies against acetyl-CoA carboxylase (ACC) (#3676, Cell Signaling Technology, MA, USA), cluster of differentiation 36 (CD36) (#28109, Cell Signaling Technology), carnitine palmitoyltransferase 1a (CPT1a) (ab234111, Abcam), fatty acid synthase (FASN) (#3180, Cell Signaling Technology), fatty acid transport protein 2 (FATP2) (#78771, Cell Signaling Technology), farnesoid X receptor (FXR) (#72105, Cell Signaling Technology), and sterol regulatory element binding protein1c (SREBP-1c) (sc-13551, Santa Cruz Biotechnology, TX, USA). .. The membranes were washed with TBST and incubated with goat anti-rabbit (#31460, Thermo Fisher Scientific, MA, USA) or goat anti-mouse secondary antibodies (#31430, Thermo Fisher Scientific) at room temperature for 1 h. The blots were incubated with β-actin (#4970, Cell Signaling Technology) as a loading control.
Binding Assay:Article Title: Tofogliflozin attenuates liver steatosis and fibrosis in non-diabetic non-alcoholic steatohepatitis mice
Article Snippet: .. The primary antibodies used in immunoblotting included antibodies against acetyl-CoA carboxylase (ACC) (#3676, Cell Signaling Technology, MA, USA), cluster of differentiation 36 (CD36) (#28109, Cell Signaling Technology), carnitine palmitoyltransferase 1a (CPT1a) (ab234111, Abcam), fatty acid synthase (FASN) (#3180, Cell Signaling Technology), fatty acid transport protein 2 (FATP2) (#78771, Cell Signaling Technology), farnesoid X receptor (FXR) (#72105, Cell Signaling Technology), and sterol regulatory element binding protein-1c (SREBP-1c) (sc-13551, Santa Cruz Biotechnology, TX, USA). .. The membranes were washed with TBST and incubated with goat anti-rabbit (#31460, Thermo Fisher Scientific, MA, USA) or goat anti-mouse secondary antibodies (#31430, Thermo Fisher Scientific) at room temperature for 1 h. The blots were incubated with β-actin (#4970, Cell Signaling Technology) as a loading control.
Article Title: Tofogliflozin attenuates liver steatosis and fibrosis in non-diabetic non-alcoholic steatohepatitis mice.
Article Snippet: .. The primary antibodies used in immunoblotting included antibodies against acetyl-CoA carboxylase (ACC) (#3676, Cell Signaling Technology, MA, USA), cluster of differentiation 36 (CD36) (#28109, Cell Signaling Technology), carnitine palmitoyltransferase 1a (CPT1a) (ab234111, Abcam), fatty acid synthase (FASN) (#3180, Cell Signaling Technology), fatty acid transport protein 2 (FATP2) (#78771, Cell Signaling Technology), farnesoid X receptor (FXR) (#72105, Cell Signaling Technology), and sterol regulatory element binding protein1c (SREBP-1c) (sc-13551, Santa Cruz Biotechnology, TX, USA). .. The membranes were washed with TBST and incubated with goat anti-rabbit (#31460, Thermo Fisher Scientific, MA, USA) or goat anti-mouse secondary antibodies (#31430, Thermo Fisher Scientific) at room temperature for 1 h. The blots were incubated with β-actin (#4970, Cell Signaling Technology) as a loading control.
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