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

Journal: Chinese Medicine

Article Title: Unraveling the potential mechanisms of Xiaochaihu decoction alleviates metabolic associated fatty liver disease (MAFLD) by integrated transcriptomics, metabolomics, and network pharmacology

doi: 10.1186/s13020-025-01310-y

Figure Lengend Snippet: XCHD regulates HFSW-induced variations of gene expression in the liver. RNA sequencing analysis was performed ( n = 3). A Volcano map of differentially expressed genes (DEGs). B Biological process (BP), cell component (CC), molecular function (MF) in GO enrichment of DEGs. C KEGG analysis of DEGs. D Pearson correlation analysis between PLINs and significantly differentially expressed lipids and lipid-like molecules. E The protein expression in liver tissue of PLIN2, PLIN3, and PLIN5 was determined by Western blot analysis. F The ratios of PLIN2/β-actin, PLIN3/β-actin, and PLIN5/β-actin are calculated ( n = 3). Data are represented using at least three independent experiments as the mean ± SD. Data are represented using at least three independent experiments as the mean ± SEM. *p < 0.05, **p < 0.01, vs. MOD group

Article Snippet: Antibodies Plin3 (10694–1-AP), Plin5 (26951–1-AP), and β-actin (81115-1-RR) were purchased from Proteintech Group (Rosemont, IL, USA).

Techniques: Gene Expression, RNA Sequencing, Expressing, Western Blot

Journal: Nutrition & metabolism

Article Title: Perilipin5 protects against non-alcoholic steatohepatitis by increasing 11-Dodecenoic acid and inhibiting the occurrence of ferroptosis.

doi: 10.1186/s12986-023-00751-2

Figure Lengend Snippet: Fig. 1 Expression of Plin5 was decreased in fatty liver models. A Venn diagram of overlapped genes in NASH group versus control group in the 4 GEO datasets. B Representation of western blot and qualification of Plin5 protein level in mice after 24 weeks of ND or HFHC feeding. C Quantification of Plin5 mRNA level in mice after 24 weeks of ND or HFHC feeding. D qPCR analysis of Plin5 mRNA level in mice under ND or CDA-HFD feeding conditions. A two-tailed Student t-test was used for statistical analysis. *< 0.05, **< 0.01. All data are shown as mean ± SEM. (n = 6/ group)

Article Snippet: Plin5 KO mice were bought and generated by Cyagen Biosciences Inc, and created using the CRISPR/ CRISPRassociated 9 (Cas9) system.

Techniques: Expressing, Control, Western Blot, Two Tailed Test

Journal: Nutrition & metabolism

Article Title: Perilipin5 protects against non-alcoholic steatohepatitis by increasing 11-Dodecenoic acid and inhibiting the occurrence of ferroptosis.

doi: 10.1186/s12986-023-00751-2

Figure Lengend Snippet: Fig. 3 Systemic knockout of Plin5 aggravated HFHC diet-induced steatohepatitis. A Schematic diagram of the experimental procedure used to examine the harmful role of Plin5 in mice fed an HFHC for 24 weeks. B–D Body weight (B), Liver mass (C), and liver mass/body weight (D) values in WT and Plin5 KO mice after feeding an HFHC diet for 24 weeks. E, F Serum TG and TC level. G Representative images of H&E and oil red O staining of liver sections from mice fed chow diet or HFHC diet. The statistics of oil red O-positive areas are shown. Scale bar, 50 um. H Relative images and quantitative data of PSR staining in liver sections from WT and Plin5 KO mice (scale bars 50 um. I, J Serum ALT (I) and AST (J) were measured in WT and Plin5 KO mice after 24 weeks of HFHC diet feeding (n = 6/group). K NAFLD activity score. L qPCR analysis of mRNA level of inflammation-related genes in the liver of WT and Plin5 KO mice fed the HFHC diet. M Relative mRNA level of profibrotic genes in liver tissue from WT and Plin5 KO mice. A two-tailed Student t test was used for statistical analysis. The data are presented as the mean ± SEM. A significant difference between the WT-ND group and the Plin5 KO-HFHC. group. *< 0.05, **< 0.01; Abbreviations: NAS, NAFLD activity score. (n = 6/group)

Article Snippet: Plin5 KO mice were bought and generated by Cyagen Biosciences Inc, and created using the CRISPR/ CRISPRassociated 9 (Cas9) system.

Techniques: Knock-Out, Staining, Activity Assay, Two Tailed Test

Journal: Nutrition & metabolism

Article Title: Perilipin5 protects against non-alcoholic steatohepatitis by increasing 11-Dodecenoic acid and inhibiting the occurrence of ferroptosis.

doi: 10.1186/s12986-023-00751-2

Figure Lengend Snippet: Fig. 4 Knockout of Plin5 diminished HFHC diet-induced ferroptosis. A, B Total and ferrous iron levels in liver were measured by iron probe in WT and Plin5 KO mice fed an HFHC-diet. C Prussian blue iron staining (Enhanced with DAB) of liver sections from mice fed an HFHC-diet. Scale bar, 50 um. D, E Serum total and ferrous iron levels were measured by iron probe in WT and Plin5 KO mice fed an HFHC-diet. F Expression of hepatic iron metabolic genes, Tfr1, Hamp1, Hamp2, Fth, Ftl, was measured by qPCR from WT and Plin5 KO mice fed an HFHC-diet. G, H MDA and SOD content was measured using a malondialdehyde assay kit (TBA method) and total superoxide dismutase assay kit (hydroxylamine method) respectively. I Hepatic mRNA level of Hmox1, Ptgs2 and Nox2 was measured by qPCR in HFHC-diet fed mice. J Confocal images of WT and Plin5 KO mouse liver sections labeled with C11-BODIPY and DAPI from mice fed on HFHC-diet. Green and blue colors indicate lipid ROS (peroxidated lipids) and nucleus respectively. Scale bar, 50 um. K Western blot analysis of GPX4 in WT and Plin5 KO mouse livers. A two-tailed Student t-test was used for statistical analysis. The data are presented as the mean ± SEM. *< 0.05, **< 0.01. Abbreviations: MDA: malondialdehyde; SOD: total superoxide dismutase. (n = 6/group)

Article Snippet: Plin5 KO mice were bought and generated by Cyagen Biosciences Inc, and created using the CRISPR/ CRISPRassociated 9 (Cas9) system.

Techniques: Knock-Out, Staining, Expressing, Labeling, Western Blot, Two Tailed Test

Journal: Nutrition & metabolism

Article Title: Perilipin5 protects against non-alcoholic steatohepatitis by increasing 11-Dodecenoic acid and inhibiting the occurrence of ferroptosis.

doi: 10.1186/s12986-023-00751-2

Figure Lengend Snippet: Fig. 5 Hepatocyte-specific overexpression of Plin5 alleviated MCD-induced NASH and ferroptosis. A Representative western blot of Plin5 measured in livers of AAV-vector and AAV-Plin5 mice. B, C Serum level of ALT and AST measured in WT and Plin5 KO mice after 3 weeks of MCD feeding. D Representative images of H&E, oil red O staining, Masson staining and PSR staining of liver sections from mice and NAFLD activity scores (Scale bar,100 um). E, F qPCR analysis of mRNA level of inflammation-related genes and profibrotic genes in the liver of AAV-vector and AAV-Plin5 mice fed an HFHC diet. G, H MDA and SOD content were measured by malondialdehyde assay kit (TBA method) and total superoxide dismutase assay kit (hydroxylamine method) respectively. I Expression of hepatic iron metabolic genes Tfr1, Hamp1, Hamp2, Fth and Ftl were measured by qPCR from AAV-vector and AAV-Plin5 mice under MCD-diet condition. J Hepatic mRNA level of Hmox1, Ptgs2, and Nox2 were measured by qPCR in MCD-diet mice. K The protein level of GPX4 and GAPDH in AAV-vector and AAV-Plin5 mice fed an MCD-diet were detected by western blot. A two-tailed Student t-test was used for statistical analysis. Data are presented as the mean ± SEM. *< 0.05, **< 0.01. (n = 6/group)

Article Snippet: Plin5 KO mice were bought and generated by Cyagen Biosciences Inc, and created using the CRISPR/ CRISPRassociated 9 (Cas9) system.

Techniques: Over Expression, Western Blot, Plasmid Preparation, Staining, Activity Assay, Expressing, Two Tailed Test

Journal: Nutrition & metabolism

Article Title: Perilipin5 protects against non-alcoholic steatohepatitis by increasing 11-Dodecenoic acid and inhibiting the occurrence of ferroptosis.

doi: 10.1186/s12986-023-00751-2

Figure Lengend Snippet: Fig. 6 Plin5 KO aggravated RSL-3 induced ferroptosis that was reversed by 11-Dodecenoic acid, a downstream targeting molecular of Plin5. A Schematic of the experimental strategy used to identify the potential target metabolites of Plin5. in steatotic hepatocytes. B Volcano plot indicates different metabolites between the Plin5 KO group and the control group. C Concentration of 11-Dodecenoic acid in WT and KO groups. D Protein expression and quantification of Plin5 in SK-HEP1 cells infected with siPlin5. E Schematic diagram of cells experiment. F Cell viability was assayed by trypan blue staining. G SK-HEP1 cells with empty vector, Plin5 knockdown, and adding 11-DA were treated with RSL-3 for 12 h and cell states were captured by microscope. Dead cells were stained by propidium iodide (PI). Scale bar, 100 um

Article Snippet: Plin5 KO mice were bought and generated by Cyagen Biosciences Inc, and created using the CRISPR/ CRISPRassociated 9 (Cas9) system.

Techniques: Control, Concentration Assay, Expressing, Infection, Staining, Plasmid Preparation, Knockdown, Microscopy