Journal: Foods

Article Title: Chemical Composition Analysis of Highland Barley ( Hordeum vulgare L.) with Different Modification Methods and Lipid Metabolism Mechanism Analysis of Highland Barley with Microwave Fluidization Modification

doi: 10.3390/foods15081396

Figure Lengend Snippet: Protein expression levels ( A , H ) including ( B ) PPARγ, ( C ) Fabp4, ( D ) CD36, ( E ) SREBP-1c, ( F ) ADIPOQ, ( G ) SCD-1, ( I ) TLR4, ( J ) IKKβ, and ( K ) p-P65 in liver tissues among NCD, HFCD, and HFCD + HB-1. Data were expressed as mean ± SD ( n = 3) based on Dunnett’s test. ** indicated p < 0.01, *** indicated p < 0.001, **** indicated p < 0.0001.

Article Snippet: The specific antibody concentrations used in this study were as follows: anti-rabbit antibodies against Peroxisome proliferator-activated receptor γ (PPARγ) (1:1000, 58 kDa, Affinity, 16643-1-AP), FABP4 (1:1000, 15 kDa, Affinity, DF6035), CD36 (1:1000, 88 kDa, Affinity, DF13262), SREBP-1c (1:1000, 122 kDa, Affinity, AF6283), ADIPOQ (1:1000, 26 kDa, Affinity, DF7000), SCD-1 (1:1000, 41 kDa, Bioss, bs-3787R), p-P65 (1:1000, 65 kDa, Affinity, AF2006, Serine Ser536), TLR4 (1:1000, 100 kDa, Affinity, AF7017), IKKβ (1:1000, 87 kDa, Affinity, AF6010), and GAPDH (1:1000, 37 kDa, Xianzhi Biotech, AB-P-R 001).

Techniques: Expressing

Journal: Cell Death and Differentiation

Article Title: SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity

doi: 10.1038/s41418-019-0356-z

Figure Lengend Snippet: SIRT3 suppresses lipogenesis in lipotoxic hepatocytes. a SCD1 protein level in vector and SIRT3OE hepatocytes treated with or without P/O mixture for 24 h. b SIRT3 and SCD1 protein levels in SIRT3 and SCD1 double overexpressed hepatocytes. The lipid content c and the cellular TG content d in SIRT3 and SCD1 double overexpressed hepatocytes. White: blank, vector; gray: blank, SIRT3OE; light gray: P/O, vector; black: P/O, SIRT3OE. Data are shown as mean ± S.D., n = 6, * p < 0.05, and *** p < 0.001, vector vs. SIRT3OE, ### p < 0.001 blank vs. P/O treatment. && p < 0.01, &&& p < 0.001, SCD1OE vs. vector. One-way ANOVA was used to calculate the p -values

Article Snippet: The plasmid of pCMV3-SCD1-N-Myc was purchased from Sino Biological (MG51311-NM, Wayne, PA, USA).

Techniques: Plasmid Preparation

Journal: Cell Death and Differentiation

Article Title: SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity

doi: 10.1038/s41418-019-0356-z

Figure Lengend Snippet: Schematic of the role of SIRT3 in protecting hepatocytes against lipotoxicity. SIRT3 activates AMPK to enhance macroautophagy and CMA on LDs and promote LDs dispersion on detyrosinated MTs, inhibits SCD1 expression to suppress lipogenesis, and deacetylates LCAD to increase fatty acid oxidation in mitochondria, resulting in attenuation of lipotoxicity in hepatocytes

Article Snippet: The plasmid of pCMV3-SCD1-N-Myc was purchased from Sino Biological (MG51311-NM, Wayne, PA, USA).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Functional Interaction Between PRDM16 and the SREBP Pathway Controls Lipid Metabolism

doi: 10.3390/ijms262110246

Figure Lengend Snippet: PRDM16 represses SREBP target gene expression and SREBP-dependent lipid metabolism. ( A ) MCF7 cells were transduced with non-targeted (C) or PRDM16 (P16) shRNA. Seventy-two hours after transduction, the media was changed to lipoprotein-deficient media, which was supplemented with 25-hydroxycholesterol (25-HC) where indicated. Ninety-six hours after transduction, mRNA was extracted and used for qPCR with primers specific for HMG-CoA synthase (HMGCS), HMG-CoA reductase (HMGCR), the LDL receptor (LDLR), fatty acid synthase (FAS), stearoyl-CoA desaturase (SCD1), and PRDM16. The relative mRNA expression in cells transduced with non-targeted shRNA and grown in the absence of 25-HC was set to 1. The data represent the average −/+ SEM of at least three independent experiments. ( B ) MCF7 cells were transduced with non-targeted (C) or PRDM16 (P16) shRNA. Seventy-two hours after transduction, whole-cell lysates were prepared and separated on SDS-PAGE gels. The amount of LDL receptor, PRDM16, and actin (loading control) was analyzed by Western blotting. ( C ) MCF7 cells were transduced as in ( B ). Seventy-two hours after transduction, the media was changed to lipoprotein-deficient media. Ninety-six hours after transduction, pHrodo-labeled LDL (red) was added to cells for 4 h, followed by repeated washes with PBS containing BSA (0.3%). The mean fluorescence intensities −/+ SD across each experimental group are provided in the bar graph (right). ( D ) MCF7 cells were transduced as in ( A ). Seventy-two hours after transduction, the media was changed to lipoprotein-deficient media alone or supplemented with 25-hydroxycholesterol (25-HC). Ninety-six hours after transduction, cells were fixed and stained with LipidTOX neutral lipid stain (green). The mean fluorescence intensities −/+ SD across each experimental group are provided in the bar graph (right). p -values lower than 0.05 were considered statistically significant. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. ns, not significant.

Article Snippet: Antibodies against PRDM16 (16212) and SCD1 (2438S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Targeted Gene Expression, Transduction, shRNA, Expressing, SDS Page, Control, Western Blot, Labeling, Fluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Functional Interaction Between PRDM16 and the SREBP Pathway Controls Lipid Metabolism

doi: 10.3390/ijms262110246

Figure Lengend Snippet: Ectopic expression of PRDM16 attenuates adipogenesis. ( A ) 3T3-L1 preadipocytes were transduced with expression vectors for GFP or PRDM16 (P16) and left untreated or differentiated to mature adipocytes (Diff). RNA was extracted, and the expression of HMG-CoA synthase (HMGCS), HMG-CoA reductase (HMGCR), the LDL receptor (LDLR), fatty acid synthase (FAS), and stearoyl-CoA desaturase (SCD1) was analyzed by qPCR. The relative expression of each gene in undifferentiated GFP-expressing cells was set to 1. The data represent the average −/+ SEM of at least three independent experiments ( B ) 3T3-L1 cells were transduced and treated as in ( A ), fixed and stained with oil red O to visualize the accumulation of lipids. The wells in the upper row are undifferentiated (−Diff), and those in the lower row are differentiated (+Diff). ( C ) 3T3-L1 cells were transduced and treated as in ( A ). RNA was extracted, and the expression of PPARγ, C/EBPα, adipsin, and SREBP1c in undifferentiated and differentiated (Diff) cells was analyzed by qPCR. The relative expression of each gene in undifferentiated GFP-expressing cells was set to 1. The data represent the average −/+ SEM of at least three independent experiments. p -values lower than 0.05 were considered statistically significant. * p < 0.05, and ** p < 0.01. ns, not significant.

Article Snippet: Antibodies against PRDM16 (16212) and SCD1 (2438S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Transduction, Staining

Journal: International Journal of Molecular Sciences

Article Title: The Functional Interaction Between PRDM16 and the SREBP Pathway Controls Lipid Metabolism

doi: 10.3390/ijms262110246

Figure Lengend Snippet: The PRDM16-mediated repression of SREBP1/2 is functional in white and brown adipocyte models. ( A ) 3T3-L1 preadipocytes were transduced with non-targeted (C) or PRDM16 (P16) shRNA. Seventy-two hours following transduction, RNA was extracted, and the expression of HMG-CoA synthase (HMGCS), HMG-CoA reductase (HMGCR), LDL receptor (LDLR), fatty acid synthase (FAS), SCD1, and PRDM16 was determined by qPCR. The relative expression of each gene in cells transduced with non-targeted shRNA was set to 1. ( B ) Human adipose-derived stem cells were transduced with non-targeted (C) or PRDM16 (P16) shRNA. Seventy-two hours following transduction, RNA was extracted, and the expression of HMG-CoA synthase ( HMGCS ), HMG-CoA reductase (HMGCR), LDL receptor (LDLR), fatty acid synthase (FAS), SCD1, and PRDM16 was determined by qPCR. The relative expression of each gene in cells transduced with non-targeted shRNA was set to 1. ( C ) 3T3-L1 preadipocytes were transduced with non-targeted (C) or PRDM16 (P16) shRNA. Ninety-six hours following transduction, cells were either left uninduced (−) or induced (+) to undergo adipocyte differentiation (Diff). Seven days after the initiation of differentiation, RNA was extracted, and the expression of C/EBPα, PPARγ, and SREBP1c was determined by qPCR. The expression of each gene in undifferentiated cells (C or P16) was set to 1. The expression of PRDM16 is shown in . ( D ) WT-1 brown preadipocytes were transduced with non-targeted (C) or PRDM16 (P16) shRNA. Ninety-six hours following transduction, RNA was extracted, and the expression of HMG-CoA synthase (HMGCS), HMG-CoA reductase (HMGCR), LDL receptor (LDLR), fatty acid synthase (FAS), SCD1, and PRDM16 was determined by qPCR. The relative expression of each gene in cells transduced with non-targeted shRNA was set to 1. The data represent the average −/+ SEM of at least three independent experiments. p -values lower than 0.05 were considered statistically significant. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Antibodies against PRDM16 (16212) and SCD1 (2438S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Functional Assay, Transduction, shRNA, Expressing, Derivative Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Glavonoid-rich oil supplementation reduces stearoyl-coenzyme A desaturase 1 expression and improves systemic metabolism in diabetic, obese KK-A y mice.

doi: 10.1016/j.biopha.2021.111714

Figure Lengend Snippet: Fig. 9. GRO supplementation reduced the protein levels of SCD1 and PPARα and increased the level of phosphorylated AMPK in the liver. (A) Western blot analysis of SCD1 in the liver of 20-week-old mice. (B) Western blot analysis of PPARα in the liver of 20-week-old mice. (C) The ratio of phosphorylated AMPK to total AMPK levels in the liver of 20-week-old mice, as determined by Western blot analysis. Cont: control mice; GRO8: 0.8% GRO mice. The data were analyzed with Student’s t- test. *p < 0.05, *p < 0.01. Values are means ± SD (n = 5).

Article Snippet: After electrophoresis, the separated proteins were transferred to a polyvinylidene difluoride membrane (Immobilon, 0.2 μm pore, (Merck Millipore Burlington, MA, USA)) and incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-mouse PPARα (1:3000, GeneTex, Los Angeles, CA, USA), β-actin (1:3000, GeneTex), total AMPKα1 (1:3000, GeneTex), phosphorylated AMPKα1 (Thr172) (1:1000, Cell Signaling Technology, Danvers, MA, USA), and SCD1 (1:1000, R&D Systems Minneapolis, MN, USA).

Techniques: Western Blot, Control

Journal: PLOS Neglected Tropical Diseases

Article Title: Single-cell sequencing analysis reveals development and differentiation trajectory of Schwann cells manipulated by M . leprae

doi: 10.1371/journal.pntd.0011477

Figure Lengend Snippet: a. Enrichment analysis of the top 100 highly expressed DEGs in cluster 3 compared with the other clusters. b. The GSEA plot of ‘PPAR signaling pathway’ and ‘peroxisome’ with higher enrichment score in cluster 3 compared with other clusters. c. Heatmap of expression of genes enriched in SREBP signalling pathway. d. Immunoblotting results show increased expression of AKR1C2 , AKR1C3 , SCD , as well as SREBP1 and SREBP2 in SCs at 72 h post-infection compared to time-matched controls.

Article Snippet: AKR1C2 antibody (PA5-36572; Invitrogen, Carlsbad, CA, USA; 1:500), AKR1C3 antibody (Abcam, ab84327; 1:500), SCD antibody (Cell Signaling Technology, #2794; 1:1000), SREBP1 antibody (2A4) (sc-13551; Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:500), SREBP2 antibody (Abcam, ab30682; 1:500), ASPM antibody (Invitrogen, PA5-99790; 1:500), TOP2A antibody (Cell Signaling Technology, #12286; 1:1000), β-ACTIN antibody (Cell Signaling Technology, #3700; 1:1000), α-TUBULIN antibody (Cell Signaling Technology, #2125; 1:1000), GAPDH antibody (60004-1-IG; Proteintech, Rosemont, IL, USA; 1:20000), horseradish peroxidase-labeled goat anti-mouse IgG (H+L) (Affinity Purified) (ZB-5305; ZSGB-BIO Co., Ltd., Beijing, China; 1:4000), and horseradish-peroxidase-labeled goat anti-rabbit IgG (H+L) (Affinity Purified) (ZSGB-BIO, ZB-5301; 1:4000) were used for western blotting.

Techniques: Expressing, Western Blot, Infection