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

Journal: bioRxiv

Article Title: Antisense oligonucleotide allele-specific targeting of EFEMP1 in a patient-derived model of Doyne honeycomb retinal dystrophy

doi: 10.1101/2025.07.16.664883

Figure Lengend Snippet: (A) IF of APOE in ISOCON, ISOKO, untreated R345W (NT) and R345W iRPE treated gymnotically with ASO1.1 (1 μM dose for 7 days from day 60 of differentiation), showing APOE (magenta) accumulation in R345W iRPE and rescue in ASO1.1 treated R345W iRPE. Nuclei are labelled with DAPI. Scale bars, 50 μm. (B) Quantitation of APOE (relative fluorescence levels) in ISOCON, untreated R345W iRPE (NT) and ASO1.1 treated R345W iRPE (1 μM dose for 7 days from day 60 of differentiation). Fluorescence levels were quantified from four independent images. Statistical significance was determined by one-way ANOVA followed by post-hoc Tukey’s (HSD) test where *, ** and *** denotes a p value <0.05, 0.01 and 0.005 respectively. Bars = mean ± SD. (C) Images of transverse sections (14 μM) from ISOCON and R345W iRPE treated gymnotically with CTRL ASO or with therapeutic ASO1.1 (1 μM and 5 μM dose for 14 days from day 84 of differentiation), showing accumulation of neutral lipid (BODIPY, green) and basal APOE (magenta) accumulation in R345W iRPE that is rescued following ASO1.1 treatment. Nuclei are labelled with DAPI. Scale bars, 20 μm.

Article Snippet: For BODIPY 493/503 (Thermo Fisher Scientific) staining, samples were incubated for 15 minutes using a 10 mM BODIPY stock that was diluted 1:1000 following 2x washes with PBS.

Techniques: Quantitation Assay, Fluorescence

Journal: bioRxiv

Article Title: Antisense oligonucleotide allele-specific targeting of EFEMP1 in a patient-derived model of Doyne honeycomb retinal dystrophy

doi: 10.1101/2025.07.16.664883

Figure Lengend Snippet: (A, B) Staining of ISOCON, untreated R345W (NT) and R345W iRPE following 2-month ASO1.1 gymnosis (1 μM) from day 100 of differentiation. (A) Accumulation of lipids (BODIPY, green) and APOE (magenta) was evident in NT R345W iRPE and was rescued following treatment with therapeutic ASO1.1. Nuclei are labelled with DAPI. Scale bars, 50 μm. (B) Accumulation of COL4 and F3 is evident in NT R345W iRPE and is rescued following treatment with therapeutic ASO1.1. Nuclei are labelled with DAPI. Scale bars, 50 μm. (C-E) Quantitation of relative fluorescence levels of BODIPY (C), APOE (D) and COL4 (E). Statistical significance was determined by one-way ANOVA followed by post-hoc Tukey’s (HSD) test where *, ** and *** denotes a p value <0.05, 0.01 and 0.005 respectively. Bars = mean ± SD. (F) TEM of ISOCON, untreated R345W (NT) and R345W iRPE following 2-month ASO1.1 gymnosis (1 μM) from day 100 of differentiation. Upper panel: ISOCON, untreated R345W and ASO1.1 treated R345W iRPE show a characteristic RPE-like morphology. Scale bars, 2μm. Lower panel: Representative TEM images of basal deposits between the basal membrane (blue outline) and the underlying transwell membrane. Scale bars, 500 nm.

Article Snippet: For BODIPY 493/503 (Thermo Fisher Scientific) staining, samples were incubated for 15 minutes using a 10 mM BODIPY stock that was diluted 1:1000 following 2x washes with PBS.

Techniques: Staining, Quantitation Assay, Fluorescence, Membrane