Journal: Molecular Metabolism

Article Title: mTORC2 and AMPK differentially regulate muscle triglyceride content via Perilipin 3

doi: 10.1016/j.molmet.2016.06.007

Figure Lengend Snippet: AMPK is sufficient for PLIN3 expression in muscle cells. (A) L6 myoblasts were incubated with 25 mM and 2.5 mM 2-deoxyglucose (2-DG) for 16 h and media lactate was determined (n = 4). (B–D) Representative western blots and quantitative analysis of p-AMPKα T172 and PLIN3 in L6 myoblasts treated as described in (A) ± Compound C (1 μM) (n = 3). (E) PLIN3 mRNA levels in L6 myotubes following 16 h of no treatment (Ctr), treatment with 2 mM AICAR (AIC), 100 μM A-769662 (AB) or both compounds (AIC + AB). Where indicated L6 cells were also treated with 10 μM Compound C (CC) (n = 4–6). (F,G) PLIN3 expression in L6 myoblasts after Ctr or AIC + AB treatment ± siRNA-mediated knockdown of FoxO1, FoxO3 or both isoforms (n = 4–6), along with (H) representative western blots with the indicated antibodies. ***p < 0.001, **p < 0.01, *p < 0.05 different from control (Ctr) or scramble (Scr); ###p < 0.001 main effect of Compound C. Values are means ± SEM.

Article Snippet: PLIN3 and β-actin probes and primers were pre-developed assay reagents from Applied Biosystems, USA (PLIN3 # mm04208646_g1 and β-actin # 4352341E).

Techniques: Expressing, Incubation, Western Blot, Knockdown, Control

Journal: Molecular Metabolism

Article Title: mTORC2 and AMPK differentially regulate muscle triglyceride content via Perilipin 3

doi: 10.1016/j.molmet.2016.06.007

Figure Lengend Snippet: mTORC2/Rictor regulates Perilipin 3 via FoxO1 in skeletal muscle. (A,B) Representative western blots and quantitative analysis of PLIN3, PLIN2 and PLIN5 in Ric WT and Ric mKO quadriceps muscle (n = 17–18). (C) Gene expression of PLIN3 in Ric WT and Ric mKO quadriceps (n = 6–7). (D) Representative western blot and quantitative analysis of nuclear FoxO1 enrichment in Ric WT and Ric mKO quadriceps (n = 12). (E) Gene expression of FoxO1 and Atrogin-1 in Ric WT and Ric mKO quadriceps (n = 7–9). (F) Representative western blot and quantitative analysis of PLIN3 in muscle-specific FoxO1 WT and FoxO1 KO muscles (n = 6). (G) Western blots with indicated antibodies in quadriceps lysates from fasted-refed Ric WT and Ric mKO mice. ***p < 0.001, **p < 0.01, *p < 0.05 are genotype differences (Ric WT vs Ric mKO) as indicated. Values are means ± SEM.

Article Snippet: PLIN3 and β-actin probes and primers were pre-developed assay reagents from Applied Biosystems, USA (PLIN3 # mm04208646_g1 and β-actin # 4352341E).

Techniques: Western Blot, Gene Expression, Muscles

Journal: Molecular Metabolism

Article Title: mTORC2 and AMPK differentially regulate muscle triglyceride content via Perilipin 3

doi: 10.1016/j.molmet.2016.06.007

Figure Lengend Snippet: AMPK is necessary for PLIN3 expression. (A,B) Representative western blots and quantitative analysis of PLIN3, PLIN2 and PLIN5 protein expression from WT quadriceps muscle or muscle over-expressing a kinase-dead AMPKα 2 construct (AMPK KD) (n = 11). (C) PLIN3 mRNA levels in wildtype and AMPK KD quadriceps muscle determined by qPCR (n = 11). (D) Representative western blot and quantitative analysis of nuclear FoxO1 protein content in AMPK wildtype and KD quadriceps (n = 9–11). (E) Triglyceride concentration in dissected quadriceps from fed AMPK wildtype and KD mice (n = 11). **p < 0.01, *p < 0.05 effects of genotype (Wildtype vs AMPK KD). Values are means ± SEM.

Article Snippet: PLIN3 and β-actin probes and primers were pre-developed assay reagents from Applied Biosystems, USA (PLIN3 # mm04208646_g1 and β-actin # 4352341E).

Techniques: Expressing, Western Blot, Construct, Concentration Assay

Journal: Molecular Metabolism

Article Title: mTORC2 and AMPK differentially regulate muscle triglyceride content via Perilipin 3

doi: 10.1016/j.molmet.2016.06.007

Figure Lengend Snippet: PLIN3 is sufficient to increase triglyceride content in muscle cells. (A) Immunoblots probing for myc-tag and GFP (n = 3) and (B) gene expression of PLIN3 (n = 4) in C2C12 myotubes 24 h after transfection with indicated constructs. (C) Dapi staining intensity relative to untreated myotubes in 24 h transfected C2C12 myotubes (n = 7). (D) Triglyceride (TG) levels in transfected myotubes treated with 2% BSA only or with 750 μM palmitate as indicated (n = 3). *p < 0.05 main effects of Myc-PLIN3 vs GFP-EV. Values are means ± SEM.

Article Snippet: PLIN3 and β-actin probes and primers were pre-developed assay reagents from Applied Biosystems, USA (PLIN3 # mm04208646_g1 and β-actin # 4352341E).

Techniques: Western Blot, Gene Expression, Transfection, Construct, Staining

Journal: Frontiers in oncology

Article Title: Proteomic Characterization of Cytoplasmic Lipid Droplets in Human Metastatic Breast Cancer Cells.

doi: 10.3389/fonc.2021.576326

Figure Lengend Snippet: FIGURE 7 | SQLE and NSDHL localize to cytoplasmic lipid droplets (CLDs) in MCF10CA1a cells. Representative immunofluorescence images of MCF10CA1a cells. Cells were stained with Alexa Fluor 633 to visualize SQLE (A) and NSDHL (B), BODIPY to visualize CLDs, and DAPI to visualize nuclei. Signals from all three channels were merged for the final image in (A, B).

Article Snippet: Cells were probed with antibodies for PLIN3, SQLE, and NSDHL (Sigma, HPA006427; SantaCruz Biotechnologies, sc-271651; Atlas Antibodies, HPA000571, respectively).

Techniques: Staining

Journal: Cell reports

Article Title: ATGL/CGI-58-Dependent Hydrolysis of a Lipid Storage Pool in Murine Enterocytes

doi: 10.1016/j.celrep.2019.07.030

Figure Lengend Snippet: (A) Radioactivity in the SI and liver of chow diet-fed WT and iDKO mice (n = 4 or 5) 30 min post-gavage of 2 μCi [9,10- 3 H(N)]-triolein in 200 μL corn oil. (B) ORO staining of duodenal sections 30 min after an oral lipid load. (C and D) Biochemical (C) and histological (D) analysis of hepatic lipid levels 30 min post-gavage of 200 μL corn oil. (E and F) Lipid concentrations (E) and (F) lipoprotein profiles in the plasma 30 min post-gavage of corn oil (200 μL). Data represent mean + SD (n = 3 or 4). *p < 0.05, **p ≤ 0.01, and ***p ≤ 0.001. Magnification, 40×; scale bar, 50 μm. (G and H) Chow diet-fed mice were fasted for 16 h prior to an oral administration of 100 μL corn oil containing 1 μg/g body weight BODIPY FL C 16 (green). Intestinal sections were co-stained with PLIN3 (red) to visualize colocalization with cLDs. PLIN3 immunofluorescence staining 30 min (G) and 2 h (H) post-gavage. Green background fluorescence results from high chlorophyll content in the chow diet (alfalfa). Arrows indicate cLDs originating from BODIPY-labeled FA, which colocalize with PLIN3; arrowheads indicate endogenous cLDs coated with PLIN3; stars indicate BODIPY-containing cLDs, which do not colocalize with PLIN3. Magnification, 100×; scale bar, 10 μm. CE, cholesteryl esters; Duo, duodenum; FC, free cholesterol; HDL, high-density lipoprotein; Hep, hepar (liver); Ile, ileum; Jej, jejunum; LDL, low-density lipoprotein; PLIN3, Perilipin 3; Sto, stomach; TC, total cholesterol; TG, triglycerides; VLDL, very low density lipoprotein. See also .

Article Snippet: Sections were incubated overnight at 4°C with the primary antibody against PLIN3 (abs482; 1:200; Merck KGaA, Darmstadt, Germany), followed by incubation with the corresponding Alexa Fluor® 594-labeled secondary antibody for PLIN3 (A11037; 1:250; Thermo Fisher Scientific, Waltham, MA).

Techniques: Radioactivity, Staining, Clinical Proteomics, Immunofluorescence, Fluorescence, Labeling

Journal: Cell reports

Article Title: ATGL/CGI-58-Dependent Hydrolysis of a Lipid Storage Pool in Murine Enterocytes

doi: 10.1016/j.celrep.2019.07.030

Figure Lengend Snippet:

Article Snippet: Sections were incubated overnight at 4°C with the primary antibody against PLIN3 (abs482; 1:200; Merck KGaA, Darmstadt, Germany), followed by incubation with the corresponding Alexa Fluor® 594-labeled secondary antibody for PLIN3 (A11037; 1:250; Thermo Fisher Scientific, Waltham, MA).

Techniques: Diagnostic Assay, Software, Imaging

Journal: Disease Markers

Article Title: Associations of Perilipin 3 with Insulin Resistance in Arab Adults with Type 2 Diabetes

doi: 10.1155/2021/4791915

Figure Lengend Snippet: Clinical characteristics of subjects according to groups.

Article Snippet: A human PLIN3 CLIA kit (Catalogue No. abx494487, Abbexa Ltd., Cambridge, UK) was used to determine PLIN3 in serum through an enzyme-linked immunosorbent assay (ELISA).

Techniques:

Journal: Disease Markers

Article Title: Associations of Perilipin 3 with Insulin Resistance in Arab Adults with Type 2 Diabetes

doi: 10.1155/2021/4791915

Figure Lengend Snippet: PLIN3 levels in the three groups expressed in terms of median, 1 st quartile (gray), and 3 rd quartile (yellow).

Article Snippet: A human PLIN3 CLIA kit (Catalogue No. abx494487, Abbexa Ltd., Cambridge, UK) was used to determine PLIN3 in serum through an enzyme-linked immunosorbent assay (ELISA).

Techniques:

Journal: Disease Markers

Article Title: Associations of Perilipin 3 with Insulin Resistance in Arab Adults with Type 2 Diabetes

doi: 10.1155/2021/4791915

Figure Lengend Snippet: Associations of PLIN3 with studied parameters according to groups.

Article Snippet: A human PLIN3 CLIA kit (Catalogue No. abx494487, Abbexa Ltd., Cambridge, UK) was used to determine PLIN3 in serum through an enzyme-linked immunosorbent assay (ELISA).

Techniques:

Journal: Disease Markers

Article Title: Associations of Perilipin 3 with Insulin Resistance in Arab Adults with Type 2 Diabetes

doi: 10.1155/2021/4791915

Figure Lengend Snippet: Significant associations of PLIN3 with (a) BMI, (b) glucose, (c) insulin, and (d) HOMA-IR.

Article Snippet: A human PLIN3 CLIA kit (Catalogue No. abx494487, Abbexa Ltd., Cambridge, UK) was used to determine PLIN3 in serum through an enzyme-linked immunosorbent assay (ELISA).

Techniques:

Journal: Disease Markers

Article Title: Associations of Perilipin 3 with Insulin Resistance in Arab Adults with Type 2 Diabetes

doi: 10.1155/2021/4791915

Figure Lengend Snippet: Independent predictors of PLIN3.

Article Snippet: A human PLIN3 CLIA kit (Catalogue No. abx494487, Abbexa Ltd., Cambridge, UK) was used to determine PLIN3 in serum through an enzyme-linked immunosorbent assay (ELISA).

Techniques:

Journal: Disease Markers

Article Title: Associations of Perilipin 3 with Insulin Resistance in Arab Adults with Type 2 Diabetes

doi: 10.1155/2021/4791915

Figure Lengend Snippet: Clinical characteristics of males and females according to studied groups.

Article Snippet: A human PLIN3 CLIA kit (Catalogue No. abx494487, Abbexa Ltd., Cambridge, UK) was used to determine PLIN3 in serum through an enzyme-linked immunosorbent assay (ELISA).

Techniques: