bodipy 493 503 staining 138 exposed zebrafish larvae  (MedChemExpress)

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