Journal: PLOS ONE

Article Title: A novel genetic model provides a unique perspective on the relationship between postexercise glycogen concentration and increases in the abundance of key metabolic proteins after acute exercise

doi: 10.1371/journal.pone.0295964

Figure Lengend Snippet: (A) Glycogen synthase abundance. (B) Glycogen. (C) HKII abundance. (D) GLUT4 abundance. (E) PGC1α abundance. (F) PDK4 abundance. (G) Representative immunoblots. (H) pAMPK Thr172 /AMPK ratio. (I) pACC1/2 Ser79/212 /ACC1/2 ratio. (J) pTBC1D1 Ser237 / TBC1D1 ratio. (K) Representative immunoblots. (L) PYGM abundance. (M) AGL abundance. (N) GBE1 abundance. (O) Representative immunoblots. shRNA-GS1 versus shRNA-Scr within the same exercise group (IPEX or Sedentary), G P < 0.05; IPEX versus Sedentary within the same AAV group (shRNA-GS1 or shRNA-Scr), E P < 0.05. Comparisons between the two groups were analyzed by a multilevel mixed-effects linear regression analysis. Values are means ± SD; n = 6/group for all the immunoblots and glycogen.

Article Snippet: Anti-glycogen synthase I (GS1; #3893), anti-hexokinase II (HKII; #2867), Anti-phospho AMPKα Thr 172 (pAMPKα Thr172 ; #50081, which recognizes phosphorylation on both α1 and α2 isoforms), anti-AMPK-α (AMPKα; #5831, which recognizes both α1 and α2 isoforms), anti-acetyl CoA carboxylase (ACC; #3676), anti-phospho ACC Ser79/212 (pACC Ser79/212 ; #3661), anti-TBC1D1 (TBC1D1; #4629), and anti-rabbit IgG horseradish peroxidase (HRP) conjugate (#7074) were from Cell Signaling Technology (Danvers, MA).

Techniques: Western Blot, shRNA

Journal: PLOS ONE

Article Title: A novel genetic model provides a unique perspective on the relationship between postexercise glycogen concentration and increases in the abundance of key metabolic proteins after acute exercise

doi: 10.1371/journal.pone.0295964

Figure Lengend Snippet: (A) Glycogen synthase abundance. (B) Glycogen. (C) HKII abundance. (D) GLUT4 abundance. (E) PGC1α abundance. (F) PDK4 abundance. (G) Representative immunoblots. (H) pAMPK Thr172 /AMPK ratio. (I) pACC1/2 Ser79/212 /ACC1/2 ratio. (J) pTBC1D1 Ser237 / TBC1D1 ratio. (K) Representative immunoblots. (L) PYGM abundance. (M) AGL abundance. (N) GBE1 abundance. (O) Representative immunoblots. shRNA-GS1 versus shRNA-Scr within the same exercise group (5hPEX or 5hSed) and diet group (refed or not refed), G P < 0.05; 5hPEX versus 5hSed within the same AAV (shRNA-GS1 or shRNA-Scr) and diet group (refed or not refed), E P < 0.05; 5h-RF versus 5h-NRF within the same AAV (shRNA-GS1 or shRNA-Scr) and exercise group (5hPEX or 5hSed), D P < 0.05. Comparisons between the two groups were analyzed by a multilevel mixed-effects linear regression analysis. Values are means ± SD; n = 6-12/group for all the immunoblots and glycogen.

Article Snippet: Anti-glycogen synthase I (GS1; #3893), anti-hexokinase II (HKII; #2867), Anti-phospho AMPKα Thr 172 (pAMPKα Thr172 ; #50081, which recognizes phosphorylation on both α1 and α2 isoforms), anti-AMPK-α (AMPKα; #5831, which recognizes both α1 and α2 isoforms), anti-acetyl CoA carboxylase (ACC; #3676), anti-phospho ACC Ser79/212 (pACC Ser79/212 ; #3661), anti-TBC1D1 (TBC1D1; #4629), and anti-rabbit IgG horseradish peroxidase (HRP) conjugate (#7074) were from Cell Signaling Technology (Danvers, MA).

Techniques: Western Blot, shRNA

Journal: PLOS ONE

Article Title: A novel genetic model provides a unique perspective on the relationship between postexercise glycogen concentration and increases in the abundance of key metabolic proteins after acute exercise

doi: 10.1371/journal.pone.0295964

Figure Lengend Snippet: (A) Glycogen synthase abundance. (B) Glycogen. (C) HKII abundance. (D) GLUT4 abundance. (E) PGC1α abundance. (F) PDK4 abundance. (G) Representative immunoblots. (H) pAMPK Thr172 /AMPK ratio. (I) pACC1/2 Ser79/212 /ACC1/2 ratio. (J) pTBC1D1 Ser237 / TBC1D1 ratio. (K) Representative immunoblots. (L) PYGM abundance. (M) AGL abundance. (N) GBE1 abundance. (O) Representative immunoblots. shRNA-GS1 versus shRNA-Scr within the same exercise group (9hPEX or 9hSed) and diet group (refed or not refed, G P < 0.05; 9hPEX versus 9hSed within the same AAV (shRNA-GS1 or shRNA-Scr) and diet group (refed or not refed), E P < 0.05; 9h-RF versus 9h-NRF within the same AAV (shRNA-GS1 or shRNA-Scr) and exercise group (9hPEX or 9hSed), D P < 0.05. Comparisons between the two groups were analyzed by a multilevel mixed-effects linear regression analysis. Values are means ± SD; n = 6/group for all the immunoblots and glycogen.

Article Snippet: Anti-glycogen synthase I (GS1; #3893), anti-hexokinase II (HKII; #2867), Anti-phospho AMPKα Thr 172 (pAMPKα Thr172 ; #50081, which recognizes phosphorylation on both α1 and α2 isoforms), anti-AMPK-α (AMPKα; #5831, which recognizes both α1 and α2 isoforms), anti-acetyl CoA carboxylase (ACC; #3676), anti-phospho ACC Ser79/212 (pACC Ser79/212 ; #3661), anti-TBC1D1 (TBC1D1; #4629), and anti-rabbit IgG horseradish peroxidase (HRP) conjugate (#7074) were from Cell Signaling Technology (Danvers, MA).

Techniques: Western Blot, shRNA

Journal: European Journal of Medical Research

Article Title: AMP-activated protein kinase α2 contributes to acute and chronic hyperuricemic nephropathy via renal urate deposition in a mouse model

doi: 10.1186/s40001-022-00800-1

Figure Lengend Snippet: AMPK α2 exacerbated kidney injury in a mouse model of acute HN. a Serum creatinine. b Blood urea nitrogen. c A pathological score of tubular injury in the kidneys. d Quantitative analysis of KIM-1-positive area in the kidneys. e Representative images of PAS staining (arrow: renal tubules dilatation, triangle: brush border shedding, square: tubular vacuolization, round: casts formation), KIM-1 immunohistochemical staining, and transmission electron microscopy. Each bar represents the mean ± SEM. Versus CON, * P < 0.05 and *** P < 0.001; versus WT, ## P < 0.01 and ### P < 0.001

Article Snippet: Anti-AMPK α2 antibody (ab3760), anti-collagen I antibody, anti-α-smooth muscle actin (α-SMA) antibody, anti-MRP4 antibody (ab180712), and anti-kidney injury molecule-1 (KIM-1) antibody were purchased from R&D systems (Minnesota, USA).

Techniques: Staining, Immunohistochemical staining, Transmission Assay, Electron Microscopy

Journal: European Journal of Medical Research

Article Title: AMP-activated protein kinase α2 contributes to acute and chronic hyperuricemic nephropathy via renal urate deposition in a mouse model

doi: 10.1186/s40001-022-00800-1

Figure Lengend Snippet: AMPK α2 exacerbated kidney injury in a mouse model of chronic HN. a Serum creatinine. b Blood urea nitrogen. c A pathological score of tubular injury in the kidneys. d Quantitative analysis of KIM-1-positive area in the kidneys. e Representative images of PAS staining (arrow: renal tubules dilatation, triangle: brush border shedding, square: tubular vacuolization, round: casts formation) and KIM-1 immunohistochemical staining. f , g Western blot analysis of KIM-1 expression in the kidneys. Each bar represents the mean ± SEM. Versus CON, *** P < 0.001; versus WT, # P < 0.05, ## P < 0.01 and ### P < 0.001

Article Snippet: Anti-AMPK α2 antibody (ab3760), anti-collagen I antibody, anti-α-smooth muscle actin (α-SMA) antibody, anti-MRP4 antibody (ab180712), and anti-kidney injury molecule-1 (KIM-1) antibody were purchased from R&D systems (Minnesota, USA).

Techniques: Staining, Immunohistochemical staining, Western Blot, Expressing

Journal: European Journal of Medical Research

Article Title: AMP-activated protein kinase α2 contributes to acute and chronic hyperuricemic nephropathy via renal urate deposition in a mouse model

doi: 10.1186/s40001-022-00800-1

Figure Lengend Snippet: AMPK α2 exacerbated renal fibrosis in a mouse model of chronic HN. a Representative images of Masson staining, Sirius red staining, α-SMA, and collagen I immunohistochemical staining. b Quantitative analysis of Masson staining-positive area in the kidneys. c Quantitative analysis of Sirius red staining-positive area in the kidneys. d Quantitative analysis of the α-SMA-positive area in the kidneys. e Quantitative analysis of collagen I-positive area in the kidneys. Each bar represents the mean ± SEM. Versus CON, * P < 0.05, ** P < 0.01 and *** P < 0.001; versus WT, # P < 0.05 and ## P < 0.01

Article Snippet: Anti-AMPK α2 antibody (ab3760), anti-collagen I antibody, anti-α-smooth muscle actin (α-SMA) antibody, anti-MRP4 antibody (ab180712), and anti-kidney injury molecule-1 (KIM-1) antibody were purchased from R&D systems (Minnesota, USA).

Techniques: Staining, Immunohistochemical staining

Journal: European Journal of Medical Research

Article Title: AMP-activated protein kinase α2 contributes to acute and chronic hyperuricemic nephropathy via renal urate deposition in a mouse model

doi: 10.1186/s40001-022-00800-1

Figure Lengend Snippet: AMPK α2 exacerbated profibrotic TGF-β/Smad3 signaling in a mouse model of chronic HN. a – e Western blot analysis of α-SMA, TGF-β1, p-Smad3 and total Smad3 expression in the kidneys. Each bar represents the mean ± SEM. Versus CON, ** P < 0.01 and *** P < 0.001; versus WT, # P < 0.05 and ## P < 0.01

Article Snippet: Anti-AMPK α2 antibody (ab3760), anti-collagen I antibody, anti-α-smooth muscle actin (α-SMA) antibody, anti-MRP4 antibody (ab180712), and anti-kidney injury molecule-1 (KIM-1) antibody were purchased from R&D systems (Minnesota, USA).

Techniques: Western Blot, Expressing

Journal: European Journal of Medical Research

Article Title: AMP-activated protein kinase α2 contributes to acute and chronic hyperuricemic nephropathy via renal urate deposition in a mouse model

doi: 10.1186/s40001-022-00800-1

Figure Lengend Snippet: AMPK α2 exacerbated renal infiltration of proinflammatory cells in a mouse model of chronic HN. a Representative immunofluorescence images of F4/80-positive macrophages in the kidneys by a confocal microscope, red color indicates positive staining of F4/80, blue color stained by DAPI indicates nuclei. b Representative images of CD3-positive T cells in the kidneys by immunohistochemical staining. c Quantitative analysis of F4/80-positive macrophages in the kidneys. d Quantitative analysis of CD3-positive T cells in the kidneys. Each bar represents the mean ± SEM. Versus CON, ** P < 0.01 and *** P < 0.001; versus WT, ## P < 0.01

Article Snippet: Anti-AMPK α2 antibody (ab3760), anti-collagen I antibody, anti-α-smooth muscle actin (α-SMA) antibody, anti-MRP4 antibody (ab180712), and anti-kidney injury molecule-1 (KIM-1) antibody were purchased from R&D systems (Minnesota, USA).

Techniques: Immunofluorescence, Microscopy, Staining, Immunohistochemical staining

Journal: European Journal of Medical Research

Article Title: AMP-activated protein kinase α2 contributes to acute and chronic hyperuricemic nephropathy via renal urate deposition in a mouse model

doi: 10.1186/s40001-022-00800-1

Figure Lengend Snippet: AMPK α2 promoted renal urate crystal deposition in a mouse model of chronic HN. a Representative images of eosin staining and compensation polarization microscopy of the entire anhydrous ethanol-treated kidneys. b Quantitative analysis of urate crystal deposition area in the kidneys. c Uric acid concentration per unit of renal tissue protein concentration. d – f Western blot analysis of AMPK α2 and MRP4 expression in the kidneys. g Genotyping of AMPK α2 knockout mice. M: DNA marker; 1, 2, wild-type homozygote, only DNA band near the 200 bp; 3, 4, heterozygote, DNA band of near both 200 bp and 600 bp; 5, 6, knockout homozygote, only DNA band near 600 bp. Each bar represents the mean ± SEM. Versus CON, * P < 0.05 and *** P < 0.001; versus WT, # P < 0.05, ## P < 0.01 and ### P < 0.001

Article Snippet: Anti-AMPK α2 antibody (ab3760), anti-collagen I antibody, anti-α-smooth muscle actin (α-SMA) antibody, anti-MRP4 antibody (ab180712), and anti-kidney injury molecule-1 (KIM-1) antibody were purchased from R&D systems (Minnesota, USA).

Techniques: Staining, Microscopy, Concentration Assay, Protein Concentration, Western Blot, Expressing, Knock-Out, Marker

Journal: European Journal of Medical Research

Article Title: AMP-activated protein kinase α2 contributes to acute and chronic hyperuricemic nephropathy via renal urate deposition in a mouse model

doi: 10.1186/s40001-022-00800-1

Figure Lengend Snippet: Schematic representation of AMPK α2 contributing to acute and chronic hyperuricemic nephropathy. AMPK α2 suppresses uric acid excretion through uric acid transporters. Accumulation of uric acid increased uric acid deposition, which further leads to the formation of a renal small tube type, inflammatory cell infiltration, and renal fibrosis

Article Snippet: Anti-AMPK α2 antibody (ab3760), anti-collagen I antibody, anti-α-smooth muscle actin (α-SMA) antibody, anti-MRP4 antibody (ab180712), and anti-kidney injury molecule-1 (KIM-1) antibody were purchased from R&D systems (Minnesota, USA).

Techniques:

Journal: International Journal of Environmental Research and Public Health

Article Title: Improved Aerobic Capacity and Adipokine Profile Together with Weight Loss Improve Glycemic Control without Changes in Skeletal Muscle GLUT-4 Gene Expression in Middle-Aged Subjects with Impaired Glucose Tolerance

doi: 10.3390/ijerph19148327

Figure Lengend Snippet: Protein expressions of AMPK- α1 ( A ) and AMPK-α2 ( B ) in skeletal muscle of IGT subjects. ( C ) Representative Western blots, using anti-AMKP-α1 and AMPK-α2 and beta-actin antibodies from skeletal muscle biopsies of non-responders and responders at baseline (0) and after the 2-year intervention.

Article Snippet: The blots were incubated overnight at +4 °C with the following antibodies: goat polyclonal anti-AMPK-α1 and -AMPK-α2 (Santa Cruz, CA, USA).

Techniques: Western Blot