Journal: Journal of Functional Foods

Article Title: Fucoidan ameliorates diabetic skeletal muscle atrophy through PI3K/Akt pathway

doi: 10.1016/j.jff.2024.106076

Figure Lengend Snippet: Fig. 7. Fucoidan ameliorates glucose metabolism in diabetic skeletal muscle and PA treated C2C12 myotubes through activating PI3K/Akt. (A) Muscle glycogen content of gastrocnemius muscle. (B) PAS staining in the gastrocnemius muscle. Magnification, ×200. Scale bar, 50 μm. (C) Muscle glycogen area in the gastroc nemius muscle. (D) Relative mRNA level of GLUT4, GSK-3β and GS. (E–F) Western blot and quantification of GLUT4, GSK-3β and GS in gastrocnemius muscle. (G) Glycogen content in C2C12 cells. (H) The different concentrations of PA treated glucose consumption of C2C12 myotubes. (I) Glucose consumption in C2C12 cells. (J) Relative mRNA level of GLUT4, GSK-3β and GS in C2C12 cells. (K–L) Western blot and quantification of GLUT4, GSK-3β and GS in gastrocnemius muscle. Data are expressed as means ± SD (n = 3). *Р < 0.05, as compared to the NC group, #Р < 0.05, as compared to the DM or PA group. &Р < 0.05, as compared to the PA + Fu group.

Article Snippet: The membrane was incubated with primary antibody for 1.5 h, including PI3K p85 (dilution 1:4000; cat# 60225-1-Ig; Proteintech, Wuhan, China), Akt (dilution 1:5000; cat# 60203-2-Ig; Proteintech, Wuhan, China), p-Akt (dilution 1:5000; cat# 60225-1-Ig; Proteintech, Wuhan, China), mTOR (dilution 1:2000; cat# ab32028; Abcam, Cambridge, MA, United States), p-mTOR (dilution 1:1000; cat# ab109268; Abcam, Cambridge, MA, United States), p70S6K (dilution 1:1000; cat# ER31205; Huabio, Hangzhou, China), p-p70S6K (dilution 1:1000; cat# 34475S; Cell Signaling Technology, Danvers, MA, United States), GLUT4 (dilution 1:2000; cat# 66846-1-Ig; Proteintech, Wuhan, China), GSK-3β (dilution 1:2000; cat# ET1607-71; Huabio, Hangzhou, China), p-GSK-3β (dilution 1:2000; cat# ET1607-60; Huabio, Hangzhou, China), GS (dilution 1:1000; cat# ET1611-59; Huabio, Hangzhou, China), FoxO1 (dilution 1:3000; cat# 18592-1-AP; Proteintech, Wuhan, China), Atrogin-1 (dilution 1:1000; cat# ET7109-25; Huabio, Hangzhou, China), MuRF1 (dilution 1:2000; cat# HA500057; Huabio, Hangzhou, China) and β-actin (dilution 1:50000; cat# ET1701-80; Huabio, Hangzhou, China).

Techniques: Staining, Western Blot

Journal: Free radical biology & medicine

Article Title: Adipocyte-secreted ANGPTL2 promotes hyperuricemia through inhibiting AKT/ABCG2 signaling.

doi: 10.1016/j.freeradbiomed.2025.03.048

Figure Lengend Snippet: Fig. 4. Angptl2 knockout improved renal UA excretion in HUA mice model (A) Angptl2 knockout and identification protocol (left panel), and genotype identification results by DNA agarose gel electrophoresis (right panel); (B) HUA mice model flow chart. (C) Fasting body weight before sacrifice. (D–G) Serum UA, CREA, BUN and XOD levels in WT, HUA, Angptl2 ko-HUA groups. (H) Urinary UA level in WT, HUA, Angptl2 ko-HUA groups. (I) Relative mRNA level of Abcg2, Slc22a8, Slc2a9 and Slc22a12 in the kidney. (J) Representative data of ABCG2 and GLUT9 in the kidney by Western blotting (left panel), and statistic data (right panel). ANGPTL2, angiopoietin-like protein 2; CREA, creatinine; BUN, blood urea nitrogen; UA, uric acid; HUA, hyperuricemia; ABCG2, ATP binding cassette subfamily G member 2; OAT3, organic anion transporter 3; GLUT9, glucose transporter 9; URAT1, urate transporter 1. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: After being deparaffinized, 0.5uM kidney sections were incubated overnight with the primary antibody ABCG2 (Abcam, USA) and GLUT9 (Proteintech, China) and stained the next day with the IHC kit (MXB Biotechnologies, China) and DAB (Proteintech, China) kit.

Techniques: Knock-Out, Agarose Gel Electrophoresis, Western Blot, Binding Assay

Journal: Free radical biology & medicine

Article Title: Adipocyte-secreted ANGPTL2 promotes hyperuricemia through inhibiting AKT/ABCG2 signaling.

doi: 10.1016/j.freeradbiomed.2025.03.048

Figure Lengend Snippet: Fig. 5. Adipocyte-specific overexpression of Angptl2 significantly increased serum UA levels in mice induced by adenine and potassium oxonate (A) HUA mice model flow chart. (B) Fasting body weight before sacrifice. (C) Serum ANGPTL2 levels after adipocyte-specific Angptl2 overexpression. (D–F) Serum CREA, BUN and UA levels after adipocyte-specific Angptl2 overexpression. (G) Relative mRNA level of Abcg2, Slc22a8, Slc2a9 and Slc22a12 in the kidney. (H) Representative data of ABCG2 and GLUT9 in the kidney by Western blotting (left panel), and statistic data (right panel). ANGPTL2, angiopoietin-like protein 2; CREA, creatinine; BUN, blood urea nitrogen; HUA, hyperuricemia; UA, uric acid; ABCG2, ATP binding cassette subfamily G member 2; OAT3, organic anion transporter 3; GLUT9, glucose transporter 9; URAT1, urate transporter 1. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: After being deparaffinized, 0.5uM kidney sections were incubated overnight with the primary antibody ABCG2 (Abcam, USA) and GLUT9 (Proteintech, China) and stained the next day with the IHC kit (MXB Biotechnologies, China) and DAB (Proteintech, China) kit.

Techniques: Over Expression, Western Blot, Binding Assay

Journal: Free radical biology & medicine

Article Title: Adipocyte-secreted ANGPTL2 promotes hyperuricemia through inhibiting AKT/ABCG2 signaling.

doi: 10.1016/j.freeradbiomed.2025.03.048

Figure Lengend Snippet: Fig. 6. ANGPTL2 decreased ABCG2 by suppressing AKT activity in HK2 cells (A) Flow chart of HK2 treatment groups. (B) UA levels in HK2 cell supernatants. (C) Relative mRNA level of Abcg2, Slc22a8, Slc2a9 and Slc22a12 in HK2 cells. (D) Representative data of ABCG2 and GLUT9 in HK2 cells by Western blotting (left panel), and statistic data (right panel). ANGPTL2, angiopoietin-like protein 2; rhA2, human recombinant proteins ANGPTL2; HUA, hyperuricemia; ABCG2, ATP binding cassette subfamily G member 2; OAT3, organic anion transporter 3; GLUT9, glucose transporter 9; URAT1, urate transporter 1. *p < 0.05, **p < 0.01, ****p < 0.0001.

Article Snippet: After being deparaffinized, 0.5uM kidney sections were incubated overnight with the primary antibody ABCG2 (Abcam, USA) and GLUT9 (Proteintech, China) and stained the next day with the IHC kit (MXB Biotechnologies, China) and DAB (Proteintech, China) kit.

Techniques: Activity Assay, Western Blot, Recombinant, Binding Assay

Journal: Free radical biology & medicine

Article Title: Adipocyte-secreted ANGPTL2 promotes hyperuricemia through inhibiting AKT/ABCG2 signaling.

doi: 10.1016/j.freeradbiomed.2025.03.048

Figure Lengend Snippet: Fig. 7. ANGPTL2 decreased ABCG2 by suppressing AKT activity in primary renal tubular epithelial cells. (A) Flow chart of RTECs treatment groups. (B) UA levels in RTECs supernatants. (C) Relative mRNA level of Abcg2, Slc22a8, Slc2a9 and Slc22a12 in RTECs. (D) Representative data of ABCG2 and GLUT9 in RTECs by Western blotting (left panel), and statistic data (right panel). ANGPTL2, angiopoietin-like protein 2; rhA2, human recombinant proteins ANGPTL2; HUA, hyperuricemia; ABCG2, ATP binding cassette subfamily G member 2; OAT3, organic anion transporter 3; GLUT9, glucose transporter 9; URAT1, urate transporter 1. *p < 0.05, **p < 0.01, ****p < 0.0001.

Article Snippet: After being deparaffinized, 0.5uM kidney sections were incubated overnight with the primary antibody ABCG2 (Abcam, USA) and GLUT9 (Proteintech, China) and stained the next day with the IHC kit (MXB Biotechnologies, China) and DAB (Proteintech, China) kit.

Techniques: Activity Assay, Western Blot, Recombinant, Binding Assay

Journal: Free radical biology & medicine

Article Title: Adipocyte-secreted ANGPTL2 promotes hyperuricemia through inhibiting AKT/ABCG2 signaling.

doi: 10.1016/j.freeradbiomed.2025.03.048

Figure Lengend Snippet: Fig. 9. Adipocyte-secreted ANGPTL2 promotes UA level through AKT/ABCG2 signaling in HK2 cells. (A) Flow diagram of C3H10 induction and treatment with FFA, as well as oil red O staining. (B) Representative data of ANGPTL2 after FFA treated C3H10 cells differentiated adipocytes by Western blotting (left panel), and statistic data (right panel). (C) ANGPTL2 levels in FFA-treated C3H10 cells differentiated adipocytes. (D) Flow chart of conditioned medium collected from C3H10 differ entiated into adipocytes treated with HK2 cells. (E) UA levels in HK2 cell supernatants after conditioned medium treatment. (F) Representative data of ABCG2 in HK2 cells by Western blotting (left panel), and statistic data (right panel). (G) Graphic summary of the study. Adipocyte-secreted ANGPTL2 decreased ABCG2 through inhibited AKT activation, then increased UA levels and led to HUA. ANGPTL2, angiopoietin-like protein 2; FFA, free fatty acid; HUA, hyperuricemia; ABCG2, ATP binding cassette G member 2; GLUT9, glucose transporter 9. *p < 0.05, **p < 0.01, ****p < 0.0001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After being deparaffinized, 0.5uM kidney sections were incubated overnight with the primary antibody ABCG2 (Abcam, USA) and GLUT9 (Proteintech, China) and stained the next day with the IHC kit (MXB Biotechnologies, China) and DAB (Proteintech, China) kit.

Techniques: Staining, Western Blot, Activation Assay, Binding Assay

Journal: Frontiers in Nutrition

Article Title: High-fat diet mouse model receiving L-glucose supplementations propagates liver injury

doi: 10.3389/fnut.2024.1469952

Figure Lengend Snippet: Metabolic profile assessment: (A) Fasting blood sugar (FBS) was assessed as indicated in materials and methods. GLUT2 expressions were quantitated by (B) western blot and (C) RT-PCR methods. (D) Serum insulin C-peptide measured by ELISA. (E) Hepatic expression of inulin receptor (IR) was evaluated from liver sections by western blot (F) HOMA2 calculator calculated HOMA-IR. Paired and unpaired Student’s t -test and ANOVA were used for statistically significant differences. p value was compared between RD and HFD control groups or within RD or HFD groups. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Rabbit anti-Human/Mouse GYS2 (Proteintech, 22371-1-AP), Rabbit anti-Human/Mouse Glycogen synthase [p Ser641] (Novus bio, NBP2-67315), rabbit anti-human/mouse PYGL antibody (Proteintech, 15851-1-AP), rabbit anti-human/mouse Glut2 polyclonal antibody (Proteintech, 20436-1-AP), rabbit anti-human/mouse ADRP/Perilipin 2 Polyclonal antibody (Proteintech, 15294-1-AP), rabbit anti-human/mouse Alpha Smooth Muscle antibody (Novus, NBP1-30894), mice anti-human/mouse AKT antibody (R&D, MAB 2055), mice anti-human/mouse phospho-AKT antibody (R&D, MAB 887), rabbit anti-human/mouse Insulin Receptor-beta antibody (Proteintech, 20433-1-AP), and rabbit anti-human/mouse beta Actin polyclonal antibody (Proteintech, 20536-1-AP).

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: Diabetes

Article Title: GLUT4 Is Not Necessary for Overload-Induced Glucose Uptake or Hypertrophic Growth in Mouse Skeletal Muscle

doi: 10.2337/db16-1075

Figure Lengend Snippet: Immunoblotting conditions

Article Snippet: Plasmids containing mouse GLUT1 (catalog #MR207871), GLUT3 (catalog #MR2097915), GLUT6 (catalog #MR219710), and GLUT10 (catalog #MR227535) and an HEK293 cell GLUT10 overexpression lysate (catalog #LY410718) were purchased from OriGene Technologies.

Techniques: Western Blot, Blocking Assay