bodipy-c16 fa  (Thermo Fisher)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Adipose Fatty Acid Binding Protein Promotes Saturated Fatty Acid-induced Macrophage Cell Death through Enhancing Ceramide Production

doi: 10.4049/jimmunol.1601403

Figure Lengend Snippet: A-FABP deficiency protects macrophages against saturated FA-induced ceramide-mediated cell death. (A) Measurement of different types of FA-induced cell death (400μM for each FA) using WT and A-FABP−/− macrophage cell lines. (B) Dual staining of 7-AAD and surface ceramide in WT and A-FABP−/− macrophage cell lines after 18h individual FA treatment (PA, SA, OA, 400μM, BSA as control). (C) WT and A-FABP−/− macrophage cell lines (0.2×106/ml/well) were cultured with designated concentrations of BODIPY-FL-C16 for 30 min, then cells were washed and harvested for flow cytometric analysis. (D, E) WT and A-FABP−/− macrophage cell lines (0.2×106/ml/well) were cultured for 16h with 100μM PA, SA, or OA on round cover slips in a 24-well plate. Cells were fixed and stained with BODIPY (green color) and DAPI (blue color) by confocal analysis (D). The fluorescence intensity of 12 typical fluorescent sites from each picture was quantified with the background subtracted in Nikon NIS elements image software (E). Experiments were performed a minimum three times (** p<0.01, ***p<0.0001 as compared to the WT control).

Article Snippet: BODIPY-C16 FA was purchased from Thermo Fisher Scientific.

Techniques: Staining, Cell Culture, Fluorescence, Software

Journal: Frontiers in Plant Science

Article Title: C1 Metabolism Inhibition and Nitrogen Deprivation Trigger Triacylglycerol Accumulation in Arabidopsis thaliana Cell Cultures and Highlight a Role of NPC in Phosphatidylcholine-to-Triacylglycerol Pathway

doi: 10.3389/fpls.2016.02014

Figure Lengend Snippet: Fluorescent PC catabolism in control, MTX-treated, and nitrogen-deprived cells. (A,C) Confocal microscopy of cells incubated for 3 days in the presence of TFPC. Two magnification are shown for both types of cells: 20 μm (upper images) and 10 μm (lower images). The fluorescence of the chromophore was measured at 503 nm following excitation at 488 nm. (B,D) Thin layer chromatography (TLC) of neutral lipids showing the effects of MTX and nitrogen starvation on PC to TAG conversion. Control, MTX-treated, and nitrogen-deprived cells, were incubated for 3 days in the presence of 20 nM TopFluor ® PC, as described in the “Materials and Methods” section. Then, cells were rinsed to remove the labeled PC before collection and lipid extraction. One hundred micrograms of lipids were deposited on the TLC and run with various standards. Fluorescence was measured at 503 nm following excitation at 488 nm. The standards Bodipy ® FL C16 (FLC16), TopFluor ® PC (TFPC), Mono-labeled DAG (DAG ∗ ), Mono-labeled TAG (TAG ∗ ) were prepared as described in the “Materials and methods” section. The cross indicates the initial deposit. (E) Distribution of the fluorescence in the different lipid species. Quantification were made from the TLC shown in (B,D) , using the ImageJ software.

Article Snippet: Fluorescent labeled PC [TopFluor ® PC, 1-palmitoyl-2-(dipyrrometheneboron difluoride)undecanoyl- sn -glycero-3-phosphocholine] was purchased from Avanti Polar Lipids, Inc. Fluorescent FA (Bodipy ® FL C16, 4,4-Difluoro-5,7-Dimethyl-4-Bora-3a,4a-Diaza-s-Indacene-3-Hexadecanoic Acid) was purchased from Thermo Fisher Scientific.

Techniques: Control, Confocal Microscopy, Incubation, Fluorescence, Thin Layer Chromatography, Labeling, Extraction, Software