Journal: Journal of Animal Science and Biotechnology

Article Title: BDH1 acetylation at K116 modulates milk fat production in dairy goats

doi: 10.1186/s40104-025-01315-5

Figure Lengend Snippet: Acetylation modification at the K116 site of BDH1 protein promotes lipid droplet accumulation and triglyceride synthesis in GMECs. A – D After transfection of pcDNA3.1-5×Flag-BDH1 (WT), K116R, or K116Q plasmids for 24 h, RT-qPCR was performed to detect the effects of acetylation/deacetylation at the K116 site of BDH1 on the expression of the following genes: ( A ) genes related to de novo fatty acid synthesis; ( B ) genes related to fatty acid desaturation and transcriptional regulation; ( C ) genes related to triglyceride synthesis, hydrolysis and lipid droplet formation; and ( D ) genes related to fatty acid activation and transport. E Lipid droplet accumulation in GMECs was observed by BODIPY staining 48 h after transfection with pcDNA3.1-5×Flag-BDH1 (wild type), K116R or K116Q plasmids. F After transfection of pcDNA3.1-5×Flag-BDH1 (wild type), K116R or K116Q plasmids for 48 h, the intracellular TAG content was determined using a commercial kit and normalized to total protein. Data were expressed as mean ± SEM, and the differences between the groups were statistically analyzed by two-tailed Student t -test or one-way analysis of variance (ANOVA), * P < 0.05, ** P < 0.01 and *** P < 0.001 indicate statistical significance

Article Snippet: After 48 h, the cells were fixed in 4% paraformaldehyde at 4 °C for 30 min, and 300 μL of BODIPY 493/503 staining solution (Invitrogen, Carlsbad, CA, USA; PBS 1:1,000 dilution) was added to each well for 30 min at room temperature, followed by 200 μL of DAPI staining solution (Beyotime, Shanghai, China) for 10 min. Lipid droplet imaging was captured by a cellular imaging reader (BioTek Instruments, Winooski, VT, USA), and lipid droplet content was expressed as BODIPY fluorescent intensity (DAPI normalized).

Techniques: Modification, Transfection, Quantitative RT-PCR, Expressing, Activation Assay, Staining, Two Tailed Test

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Pharmacological induction of acetyl-CoA carboxylase 1 autophagic degradation attenuates lipid accumulation and cholangiocarcinoma progression

doi: 10.1186/s13046-025-03564-8

Figure Lengend Snippet: WA significantly attenuates lipid accumulation in CCA. A UMAP visualization of integrated scRNA-seq data (20 tumors, 12 adjacent tissues) showing seven major cell lineages, and malignant cholangiocyte identification by CopyKat analysis (dark blue: malignant, yellow: non-malignant). B Metabolic pathway enrichment in malignant versus non-malignant cholangiocytes. C Single-cell flux estimation analysis (scFEA) was performed to infer metabolic fluxes at single-cell resolution, revealing differential metabolite profiles and metabolic activities between malignant and non-malignant cholangiocytes. D CK19 IHC staining and oil red O staining to detect lipid accumulation in human CCA tissues, scale bar = 20 μm. E CK19 IHC staining and oil red O staining to detect lipid accumulation in the KRAS/P19 CCA model, scale bar = 20 μm. F CK19 IHC staining and oil red O staining to detect lipid accumulation in the YAP/AKT CCA model, scale bar = 20 μm. G The schematic for screening of anti-tumor inhibitors targeting lipid accumulation in a natural compound library containing 994 compounds. H The inhibitory rate of each inhibitor on HuCCT1 and RBE was detected by CCK8 assay. Red coloration indicated inhibitors that have achieved more than 50% inhibition. I BODIPY493/503 fluorescence staining and flow cytometry experiments were performed to detect lipid droplet content in HuCCT1 and RBE after treatment with each of the 24 compounds. J Molecular structure of WA. K This study employed a combination of BODIPY 493/503 fluorescence staining and Oil Red O staining to detect lipid droplet content in RBE and HuCCT1 cells 24 h after WA treatment. scale bar = 100 μm. L - M Lipids in WA treatment CCA cells were measured by LC-MS. The heatmap highlighted relative changes for each type of lipid metabolite with their individual average ion counts normalized to NTC. Data are representative of three independent experiments. scale bar = 100 μm. The data are represented as the mean ± SD of three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, no significance

Article Snippet: For BODIPY 493/503 staining: 100 μL of 10 μM BODIPY 493/503 working solution (MCE, NJ, USA) was added to each well and incubated for 30 min.

Techniques: Immunohistochemistry, Staining, Drug discovery, CCK-8 Assay, Inhibition, Fluorescence, Flow Cytometry, Liquid Chromatography with Mass Spectroscopy

Journal: Advanced Science

Article Title: Microglial Feimin Alleviates Cognitive Impairment in High‐Fat Diet‐Fed Mice

doi: 10.1002/advs.202512023

Figure Lengend Snippet: Inhibition of the AKT pathway relieves palmitic acid (PA)‐induced lipid droplet (LD) accumulation and inflammation in feimin‐specific knockdown BV2 cells. A) Feimin‐specific knockdown BV2 cells followed by PA treatment for 24 h and B) The primary microglia from the microglial feimin conditional knockout mice (feimin Mic‐/− ) after 12 weeks of high‐fat diet (HFD) were pretreated with DMSO or MK2206 (AKT inhibitor, 10 µM) for 30 min, cells were then immunostained for PLIN2 + /BODIPY + (LDs). N = 5, n = 4 biological replicates. Scale bar: 50 µm. C,D) The percentage of cells displaying BODIPY + feimin + and PLIN2 + feimin + puncta was quantified, n = 5, p = 0.007 and 0.0006, p = 0.041and 0.006, n = 4 biological replicates. E–H) IL‐1β and IL‐6 mRNA (E, F, p = 0.0005, **** p < 0.001) and protein (G, H, p = 0.0006 and 0.0032) in feimin siRNA‐treated BV2 cells pretreated with DMSO or MK2206 for 30 min, followed by PA treatment for 24 h, were determined by qPCR and ELISA, respectively. I–L) IL‐1β and IL‐6 mRNA (I, J, p = 0.0006 and 0.0004,) and protein (K, L, p = 0.033 and 0.0005) in feimin Mic‐/− primary microglia pretreated with DMSO or MK2206 for 30 min, were determined by qPCR (n = 3) and ELISA (n = 5), respectively, n = 3 biological replicates. (A–L) Data were presented as means ± SD. Two‐tail Student's T‐test.

Article Snippet: To visualize LDs in brain tissues, sections were washed and incubated with BODIPY 493/50 solution (1:1000 in PBS; Thermo Fisher Scientific, D3922) for 15 min at RT.

Techniques: Inhibition, Knockdown, Knock-Out, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Microglial Feimin Alleviates Cognitive Impairment in High‐Fat Diet‐Fed Mice

doi: 10.1002/advs.202512023

Figure Lengend Snippet: High‐fat diets (HFDs) induce feimin expression, whereas microglial feimin knockout enhances HFD‐induced lipid droplet (LD) accumulation. A) Immunostaining of feimin and IBA1+ (microglia) in the hippocampal CA3 region of control (standard diet [SD]) and HFD mice. Yellow arrow indicates feimin + microglia. n = 5 mice per group, n = 3 biological replicates. Scale bar: 50 µm. B) The percentages of feimin + in IBA1 + microglia ( p = 0.027) and C) the numbers of Iba1 + microglia /mm 2 were shown ( p = 0.002). n = 5 mice per group. D) Feimin mRNA expression was determined by qPCR in the hippocampus of the control and HFD groups (n = 5, p = 0.0003). E) Immunostaining of BODIPY + (LDs) and IBA1 + (microglia). Representative LDs are represented by yellow arrows. Scale bar: 50 µm. n = 6 mice per group. The percentages of BODIPY + in IBA1 + microglia from the hippocampi CA1 (F) (n = 6, p = 0.002 and 0.031), CA3 G) (n = 6, p = 0.023 and 0.0004), and DG H) (n = 6, p = 0.004 and 0.003) region from SD+feimin fl/fl , SD+feimin Mic‐/− , HFD + feimin fl/fl , and HFD + feimin Mic‐/‐ mice. (A–H) Data were presented as means ± SD of three independent experiments. (B‐D) Two‐tail Student's T‐test, (F–H) One‐Way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: To visualize LDs in brain tissues, sections were washed and incubated with BODIPY 493/50 solution (1:1000 in PBS; Thermo Fisher Scientific, D3922) for 15 min at RT.

Techniques: Expressing, Knock-Out, Immunostaining, Control