Journal: Research

Article Title: EVA1A Regulates Hepatic Lipid Homeostasis by Modulating CD36 Expression and Its Palmitoylation

doi: 10.34133/research.1001

Figure Lengend Snippet: EVA1A deletion promotes the palmitoylation of CD36. (A to C) The palmitoylation levels of CD36 were assessed using the APE assay after treatment with 2-BP (50 μM, 6 h) as indicated, with or without HA. The top band indicated the palmitoylated CD36 (PEG-CD36). Detection of palmitoylated CD36 in EVA1A-knockdown HepG2 and Huh7 cells, along with control cells (A). Detection of palmitoylated CD36 in EVA1A-overexpressed HepG2 and Huh7 cells or control cells (B). Detection of palmitoylated CD36 in live tissues of Eva1a +/+ mice and Eva1a −/− mice ( n = 3) (C). Protein levels of palmitoylated CD36 were quantified in the right panels. (D to G) Relative mRNA levels of genes related to palmitoyl transferases (Z DHHC4 and Z DHHC5 ) and acyl-protein thioesterase 1 ( LYPLA1 ) in vitro (D and E) and in vivo (F and G). (H) Western blot analysis of ZDHHC5, ZDHHC4, and APT1 in EVA1A-knockdown or overexpressed HepG2 and Huh7 cells, after 6-h treatment with 400 μM OA or after being left untreated. (I) Western blot analysis of ZDHHC5, ZDHHC4, and APT1 in liver tissues of Eva1a +/+ mice and Eva1a −/− mice ( n = 3). Their protein levels were quantified in the lower panels. * P < 0.05, **P < 0.01, ***P < 0.001. HA: NH 2 OH.

Article Snippet: For cell treatment, the final concentrations of OA (Sangon, China), 2-BP (MCE, USA), Torin-1 (Aladdin, China), and GW9662 (Aladdin, China) were 400 μM, 50 μM, 300 nM, and 5 μM, respectively.

Techniques: Knockdown, Control, In Vitro, In Vivo, Western Blot

Journal: Research

Article Title: EVA1A Regulates Hepatic Lipid Homeostasis by Modulating CD36 Expression and Its Palmitoylation

doi: 10.34133/research.1001

Figure Lengend Snippet: Inhibition of CD36 palmitoylation attenuates EVA1A deficiency-induced fatty acid uptake and lipid droplet accumulation. (A and E) The EVA1A-knockdown HepG2 or Huh7 cells were transfected with empty vector (pcDNA3.1) or wt-CD36, or mut-CD36 plasmid for 24 h, which had been treated with 400 μM OA during the final 6 h, then were fixed and stained with BODIPY FL-C16 (A) or ORO (E). Scale bars: 20 μm. The fluorescence signal intensity of FL-C16 per cell was quantified by ImageJ. Values were means ± SDs ( n = 50). (B) Cellular NEFA levels in cells subjected to the treatment shown in (A). (C and G) The EVA1A-knockdown cells were co-treated with 50 μM 2-BP and 400 μM OA for 6 h, then were subjected to BODIPY FL-C16 staining (C) or ORO staining (G). Scale bars: 20 μm. (D) Cellular NEFA levels in cells subjected to the treatment shown in (C). (F) Cellular TG contents in cells treated as described in (A). (H) Cellular TG contents in cells treated as described in (C). * P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: For cell treatment, the final concentrations of OA (Sangon, China), 2-BP (MCE, USA), Torin-1 (Aladdin, China), and GW9662 (Aladdin, China) were 400 μM, 50 μM, 300 nM, and 5 μM, respectively.

Techniques: Inhibition, Knockdown, Transfection, Plasmid Preparation, Staining, Fluorescence

Journal: Frontiers in Neuroscience

Article Title: Revealing cerebrospinal fluid biomarkers in Parkinson's disease dementia based on iTRAQ proteomics research

doi: 10.3389/fnins.2025.1682274

Figure Lengend Snippet: PDD patients had decreased CSF IGFBP3 level compared to PD patients. ELISA results of IGFBP3 detection showed that IGFBP3 concentration was lower in the CSF of PDD patients than PD patients ( n = 28 in PD group and n = 9 in PDD group). Columns with error bars present the mean ± SEM. Two-tailed unpaired t -tests. * p < 0.05.

Article Snippet: Concentrations of IGFBP3, NXPH1, LRRN1, and HPRT1 in CSF were measured using human ELISA kits (IGFBP3: CSB-E04590h, CUSABIO, China; NXPH1: ELK3869, ELK Biotechnology, China; LRRN1: MBS2602720, MyBioSource, USA; HPRT1: ELK3546, ELK Biotechnology, China).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Two Tailed Test