Journal: Foods (Basel, Switzerland)

Article Title: Hericium erinaceus Protein Alleviates High-Fat Diet-Induced Hepatic Lipid Accumulation and Oxidative Stress In Vivo.

doi: 10.3390/foods14030459

Figure Lengend Snippet: Figure 6. Effect of HEP on the mRNA expression of hepatic lipid metabolism-related genes in obese mice: (A) PPARα (fold); (B) LPL (fold); (C) CPT-1a (fold); (D) ACOX1 (fold); (E) SREBP-1c (fold); (F) ACC1 (fold); (G) SCD-1 (fold); and (H) FASN (fold). The different lowercase letters (a, b, c, d) indicate significant differences between values in different groups (p < 0.05).

Article Snippet: Primary antibodies PPARα, CPT-1a, ACOX1, SREBP-1c, SCD-1, and FASN were purchased from Proteintech (Wuhan, China).

Techniques: Expressing

Journal: Foods (Basel, Switzerland)

Article Title: Hericium erinaceus Protein Alleviates High-Fat Diet-Induced Hepatic Lipid Accumulation and Oxidative Stress In Vivo.

doi: 10.3390/foods14030459

Figure Lengend Snippet: Figure 7. Effects of HEP on the protein expression of hepatic lipid metabolism-related genes in obese mice: (A) Representative images of Western blot; (B) PPARα (fold); (C) CPT-1a (fold); (D) ACOX1 (fold); (E) SREBP-1c (fold); (F) SCD-1 (fold); and (G) FASN (fold). The different lowercase letters (a, b, c, d) indicate significant differences between values in different groups (p < 0.05).

Article Snippet: Primary antibodies PPARα, CPT-1a, ACOX1, SREBP-1c, SCD-1, and FASN were purchased from Proteintech (Wuhan, China).

Techniques: Expressing, Western Blot

Journal: The Journal of Endocrinology

Article Title: DHT causes liver steatosis via transcriptional regulation of SCAP in normal weight female mice

doi: 10.1530/JOE-21-0040

Figure Lengend Snippet: qRT-PCR primer sequences.

Article Snippet: AR was transiently transfected as done before ( Han et al 2019 ) using Lipofectamine 2000 (Thermo Fisher) with a pSV Sport SREBP-1c vector containing an SREBP1c overexpression vector or a pSV-Sport empty vector (Addgene, Cambridge, MA).

Techniques: Binding Assay

Journal: The Journal of Endocrinology

Article Title: DHT causes liver steatosis via transcriptional regulation of SCAP in normal weight female mice

doi: 10.1530/JOE-21-0040

Figure Lengend Snippet: Proposed model: obesity-independent NAFLD in normal weight PCOS-like mouse model. (A) Here we show two proposed mechanisms by which DHT may increase hepatic lipid content. The steps are as follows: In the liver of control mice (normal physiology), SREBP1 remains in the ER via binding with SCAP and INSIG. When sterol is low (in the fed state of normal physiology ), SCAP dissociates from INSIG and the SCAP-SREBP1 complex is translocated to the Golgi. Then, proteases, S1P and S2P, cleave SREBP1, releasing the active version of SREBP1 into the cytosol. The active SREBP1 travels to the nucleus, where it activates genes involved in lipid synthesis. In the liver of DHT mice , (1) DHT enters the cell, binds, and activates AR, (2) AR enters the nucleus and binds to intron 8 of SCAP leading to increased Scap mRNA and subsequently increased SCAP protein levels, (3) increased SCAP protein expression and increased SCAP binding to the 65 kD cleaved inactive SREBP1 leads to, (4) increased nuclear (active) SREBP1 which leads to, (5) increased lipogenic mRNA expression resulting in increased de novo lipogenesis. Overall, AR binding to intron 8 in Scap may be a mechanism leading to increased SCAP which led to SCAP-SREBP1 binding being elevated which led to increased active nuclear SREBP1 and increased lipogenic gene expression. Key: Larger green arrows indicate greater increase as compared to the thinner black arrows. The red box highlights the proposed mechanism. (B) Transcriptional control of lipogenesis and lipid metabolism in the liver: de novo lipogenesis (DNL) is known to be controlled by glucose and insulin signaling pathways, leading to increased expression of lipogenic genes. What is known : (1) Insulin stimulates the activity of SREBP1c, a transcription factor that augments lipogenic enzymes (ACC1, FAS, SCD1), via activation of LXR and several other methods not depicted (for more details see Dorotea et al. 2020 ). FXR lowers lipid synthesis by decreasing SREBP1c and LXRα activity, (2) glucose promotes the activity of ChREBP, another transcription factor that increases lipogenic and glycolytic (not depicted) enzymes. ChREBP is directly regulated by LXRs as well and it controls the amount of MUFA-to-SFA, creating more MUFA by activating SCD1. Glucose has also been shown to activate LXR’s genes, (3) free fatty acids (FFAs) from the serum enter the hepatocytes from fatty acid transport protein 2 (FATP2) and are incorporated into lipid droplets with the assistance of PPARγ which stimulates FA storage . What we discovered here : (4) We examined the effect of low-dose DHT on all of the genes and proteins depicted in Fig. 5B and none of them were altered by DHT except SCAP, SREBP1, FAS, and ACC. As depicted in Fig. 5A, low-dose DHT increased SCAP mRNA, protein, and binding to SREBP1 leading to increased FAS and ACC leading to increased hepatic lipid content. ACC, acetyl-CoA carboxylase; ChREBP, carbohydrate-responsive element-binding protein; FA, fatty acid; FAS, fatty acid synthase; FFAs, free fatty acids; FxR, farnesoid X receptor; LxR, liver X receptor; MUFA, monosaturated fatty acids; PPARγ, peroxisomal proliferator-activated receptor gamma; SCAP, SREBP cleavage-activating protein; SCD1, stearoyl CoA desaturase 1; SFA, saturated fatty acids; SREBP1c, sterol regulatory element-binding protein 1c; TG, triglyceride.

Article Snippet: AR was transiently transfected as done before ( Han et al 2019 ) using Lipofectamine 2000 (Thermo Fisher) with a pSV Sport SREBP-1c vector containing an SREBP1c overexpression vector or a pSV-Sport empty vector (Addgene, Cambridge, MA).

Techniques: Binding Assay, Expressing, Activity Assay, Activation Assay

Journal: Scientific reports

Article Title: High throughput compound screening in neuronal cells identifies statins as activators of ataxin 3 expression.

doi: 10.1038/s41598-023-41192-4

Figure Lengend Snippet: Figure 2. Simvastatin treatment increases ATXN3 mRNA and protein levels, induces HMGCR protein level and activates the SREBP1 transcription factor and leads to binding of activated SREBP1 to the ATXN3 promotor. (A) qRT-PCR of Simvastatin treated SK-N-SH wild type cells for 1–2 h. Data were normalized to three house—keeping genes and DMSO controls. (B) Relative n-fold enrichment of ATXN3 promotor of three independent ChIP-qPCR experiments with the SREBP1 antibody (Proteintech 14088-1-AP) after 1 h Simvastatin (Sim) treatment. (C) Representative Western Blot (WB) of Sim treatment for 2–8 h. (D) Relative n-fold change of protein levels for mSREBP1, (E) ATXN3 and (F) HMGCR. Asterix and arrow indicate transcriptionally inactive- and mSREBP1, respectively. Hash: unspecific band. qRT-PCR Data were analysed using the ΔΔCT method by Pfaffl. ChIP-qPCR data were normalized to input and DMSO controls and represent the mean of three technical replicates for each experiment, respectively. WB data were normalized to actin and the mean of DMSO controls. Error bars indicate SD of three biological repetitions. For all experiments, a final concentration of 10 µM Sim was applied. Adjusted p values: *p < 0.05; **p = 0.005; ***p = 0.0002; ****p < 0.0001.

Article Snippet: Simvastatin treatment increases ATXN3 mRNA and protein levels, induces HMGCR protein level and activates the SREBP1 transcription factor and leads to binding of activated SREBP1 to the ATXN3 promotor. (A) qRT-PCR of Simvastatin treated SK-N-SH wild type cells for 1–2 h. Data were normalized to three house—keeping genes and DMSO controls. (B) Relative n-fold enrichment of ATXN3 promotor of three independent ChIP-qPCR experiments with the SREBP1 antibody (Proteintech 14088-1-AP) after 1 h Simvastatin (Sim) treatment. (C) Representative Western Blot (WB) of Sim treatment for 2–8 h. (D) Relative n-fold change of protein levels for mSREBP1, (E) ATXN3 and (F) HMGCR.

Techniques: Binding Assay, Quantitative RT-PCR, ChIP-qPCR, Western Blot, Concentration Assay

Journal: Marine Drugs

Article Title: Effect of Marine Microalga Chlorella pyrenoidosa Ethanol Extract on Lipid Metabolism and Gut Microbiota Composition in High-Fat Diet-Fed Rats

doi: 10.3390/md16120498

Figure Lengend Snippet: Effect of CPE55 on the mRNA and protein expressions levels in the liver. The mRNA expression ( A ) and protein expression ( B , C ) of Acetyl CoA carboxylase (ACC), AMPK-α, 3-Hydroxy-3-methyl glutaryl coenzyme A reductase (HMG-CoA), and Sterol regulatory element-binding transcription factor-1c (SREBP-1c) levels were determined through real-time quantitative PCR (RT-qPCR) and western blotting analysis. Data are expressed as the mean ± SD. One-way ANOVA with Tukey’s test. * p < 0.05 and ** p < 0.01 for CPE55 versus NFD; # p < 0.05, ## p < 0.01 for CPE55 versus HFD.

Article Snippet: The membranes were incubated for 3.5 h at 37 °C using rabbit polyclonal antibodies against GAPDH, HMG-CoA, SREBP-1c, ACC, and AMPK-α (1:1000; Beyotime Biotechnology, Shanghai, China).

Techniques: Expressing, Binding Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot