Journal: Scientific Reports

Article Title: Perilipin 5 fine-tunes lipid oxidation to metabolic demand and protects against lipotoxicity in skeletal muscle

doi: 10.1038/srep38310

Figure Lengend Snippet: Representative blots ( A ) and quantification of PLIN5 ( B ) and ATGL ( C ) protein content in different mouse skeletal muscles (n = 5) (EDL: extensor digitorum longus , TA: tibialis anterior , Sol: soleus ). **p < 0.01, ***p < 0.001 versus EDL. ( D ) Quantification of PLIN5 protein content in vastus lateralis muscle of healthy lean and endurance-trained volunteers (n = 11 per group). Correlations between muscle PLIN5 protein and ( E ) cytochrome oxidase activity, and ( F ) glucose disposal rate (n = 33). *p < 0.05 versus lean.

Article Snippet: For overexpression experiments, adenoviruses expressing in tandem GFP and human PLIN5 (hPLIN5) were used (Vector Biolabs, Philadelphia, PA).

Techniques: Muscles, Activity Assay

Journal: Scientific Reports

Article Title: Perilipin 5 fine-tunes lipid oxidation to metabolic demand and protects against lipotoxicity in skeletal muscle

doi: 10.1038/srep38310

Figure Lengend Snippet: ( A ) Representative blot and quantification of PLIN5 protein content in control (Ad-GFP) and PLIN5-overexpressing myotubes (Ad-PLIN5) (n = 3). Pulse-Chase studies using [1- 14 C] oleate were performed to determine ( B ) FA release into the culture medium (Ad-GFP = 59 ± 1.8 nmol/3 h/mg protein), ( C ) FA oxidation (Ad-GFP = 2.22 ± 0.13 nmol/3 h/mg protein), and the rate of incorporation of radiolabeled oleate into ( D ) TAG, ( E ) DAG (T0 Ad-GFP = 3.14 ± 0.14 nmol/3 h/mg protein) and ( F ) intracellular FA content (T0 Ad-GFP = 0.42 ± 0.03 nmol/3 h/mg protein) in control (Ad-GFP) and PLIN5-overexpressing myotubes (Ad-PLIN5). ( G ) Glycogen synthesis and ( H ) glucose oxidation were measured in control myotubes (Ad-GFP) and myotubes overexpressing PLIN5 (Ad-PLIN5) using [U- 14 C] glucose. ( I ) PDK4 gene expression was measured in control (Ad-GFP) and PLIN5-overexpressing myotubes (Ad-PLIN5). (n = 6) *p < 0.05, **p < 0.01 ***p < 0.001 versus Ad-GFP.

Article Snippet: For overexpression experiments, adenoviruses expressing in tandem GFP and human PLIN5 (hPLIN5) were used (Vector Biolabs, Philadelphia, PA).

Techniques: Control, Pulse Chase, Gene Expression

Journal: Scientific Reports

Article Title: Perilipin 5 fine-tunes lipid oxidation to metabolic demand and protects against lipotoxicity in skeletal muscle

doi: 10.1038/srep38310

Figure Lengend Snippet: Pulse-Chase studies using [1- 14 C] oleate were performed to determine the rate of ( A,D ) incorporation of radiolabeled oleate into TAG and ( B,E ) oleate oxidation in control myotubes (Ad-GFP) and myotubes overexpressing PLIN5 (Ad-PLIN5) either during ( A–C ) forskolin (FK) (Ad-GFP CONT = 1.66 ± 0.25 nmol/3 h/mg protein) or ( D–E ) electrical pulse (EPS) stimulation (Ad-GFP CONT = 6.33 ± 2.05 nmol/24 h/mg protein) (n = 6). Values are expressed in % of Ad-GFP Control ( A,B,D,E ) and in fold change over control in ( C and F ). *p < 0.05, **p < 0.01 ***p < 0.001 versus Ad-GFP.

Article Snippet: For overexpression experiments, adenoviruses expressing in tandem GFP and human PLIN5 (hPLIN5) were used (Vector Biolabs, Philadelphia, PA).

Techniques: Pulse Chase, Control

Journal: Scientific Reports

Article Title: Perilipin 5 fine-tunes lipid oxidation to metabolic demand and protects against lipotoxicity in skeletal muscle

doi: 10.1038/srep38310

Figure Lengend Snippet: ( A ) Glycogen synthesis was measured in control myotubes (Ad-GFP) and myotubes overexpressing PLIN5 (Ad-PLIN5) using [U- 14 C] glucose in absence or presence of 100 nM insulin, in control cells and in cells treated with 300 μM of palmitic acid for 24 h (n = 9). ( B ) Total diacylglycerols (DAG) and ( C ) Ceramide (CER) d18:1/16:0 content were measured in control myotubes (Ad-GFP) and myotubes overexpressing PLIN5 (Ad-PLIN5) (n = 4). *p < 0.05, **p < 0.01 versus Ad-GFP.

Article Snippet: For overexpression experiments, adenoviruses expressing in tandem GFP and human PLIN5 (hPLIN5) were used (Vector Biolabs, Philadelphia, PA).

Techniques: Control

Journal: Scientific Reports

Article Title: Perilipin 5 fine-tunes lipid oxidation to metabolic demand and protects against lipotoxicity in skeletal muscle

doi: 10.1038/srep38310

Figure Lengend Snippet: PLIN5 ( A ) gene expression and ( B ) protein content measured in control (shNT) and PLIN5 silenced (shPLIN5) mouse tibialis anterior muscle (n = 6). Palmitate ( C ) and glucose ( D ) oxidation rate were measured using respectively [U- 14 C] glucose or [1- 14 C] palmitate in control (shNT) and PLIN5 silenced (shPLIN5) muscle homogenates. Palmitate oxidation (i.e. CO2), acid soluble metabolites accumulation (i.e. ASMs) and total oxidation (i.e. the sum of CO2 release and ASMs accumulation) were measured (n = 6). ( E ) Insulin-stimulated glucose uptake was determined in control (shNT) and PLIN5 knockdown (shPLIN5) muscles. (n = 7). *p < 0.05, ***p < 0.001 versus shNT.

Article Snippet: For overexpression experiments, adenoviruses expressing in tandem GFP and human PLIN5 (hPLIN5) were used (Vector Biolabs, Philadelphia, PA).

Techniques: Gene Expression, Control, Knockdown, Muscles

Journal: Scientific Reports

Article Title: Perilipin 5 fine-tunes lipid oxidation to metabolic demand and protects against lipotoxicity in skeletal muscle

doi: 10.1038/srep38310

Figure Lengend Snippet: PLIN5 ( A ) gene expression and ( B ) protein content measured in control (shNT) and PLIN5 silenced (shPLIN5) mouse tibialis anterior muscle (n = 6). ( C ) Insulin-stimulated glucose uptake, ( D ) total ceramide (CER) and ( E ) total diacylglycerols (DAG) content were determined in control (shNT) and PLIN5 knockdown (shPLIN5) muscles. (n = 7). ( F ) Insulin-stimulated Akt phosphorylation on Ser473 and Thr308 residues was measured in control (shNT) and PLIN5 silenced (shPLIN5) muscle (n = 4). *p < 0.05, **p < 0.01, ***p < 0.001 versus shNT.

Article Snippet: For overexpression experiments, adenoviruses expressing in tandem GFP and human PLIN5 (hPLIN5) were used (Vector Biolabs, Philadelphia, PA).

Techniques: Gene Expression, Control, Knockdown, Muscles, Phospho-proteomics

Journal: Scientific Reports

Article Title: Perilipin 5 fine-tunes lipid oxidation to metabolic demand and protects against lipotoxicity in skeletal muscle

doi: 10.1038/srep38310

Figure Lengend Snippet: In the resting state, PLIN5 protects LD from lipolytic attack by lipases. An increase in PLIN5 content (red arrows) slows down lipolysis and FA oxidation, favoring a switch towards glucose utilization. During lipolytic stimulation (i.e. PKA activation or contraction), PLIN5 enhances FA oxidation, thereby increasing CO 2 production. It has been suggested that PLIN5 could provide a physical linkage between LD and mitochondria. We can hypothesize that this relocation has metabolic consequences by facilitating FA channeling from LD to mitochondria, thus allowing a more efficient coupling between IMTG lipolysis and FA oxidation upon increased metabolic demand. Finally, the up-regulation of PLIN5 with high-fat feeding is insufficient to protect from LD-mediated CER accumulation. FA: Fatty Acids; IMTG: Intramyocellular Triacylglycerols; CER: Ceramides.

Article Snippet: For overexpression experiments, adenoviruses expressing in tandem GFP and human PLIN5 (hPLIN5) were used (Vector Biolabs, Philadelphia, PA).

Techniques: Activation Assay