SNB19 cells were grown in 4-well chamber slides either with 10% FBS DMEM or 10% lipid-depleted FBS (Biowest) DMEM for 48 hours. BTSC cells were grown in 4-well chamber slides in complete Neurobasal medium, as previously described. For lipid starvation SNB19 cells were kept for additional 2 hours either in 10% lipid-depleted FBS DMEM or in 10% FBS DMEM. All cells were then fixed with 4% paraformaldehyde in PBS and processed for the staining. FLAG-ZBTB18 (FL or Nte) was labeled with anti-FLAG M2 monoclonal primary antibody (Sigma #F1804). CTBP2 was labeled with anti-CTBP2 mouse monoclonal antibody (BD Biosciences # 612044). Lipid droplets were stained with anti-Perilipin-3 rabbit polyclonal antibody (Abcam #ab47638) (SNB19 cells), or with 0.5μg/ml Bodipy TMR-X SE (Thermo Fisher Scientific, #D6117) in 150mM NaCl for 10 minutes at room temperature (BTSC475). Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich). Pictures were acquired using a FSL confocal microscope (Olympus).
Bodipy tmr x se
Manufactured by Thermo Fisher Scientific
Bodipy TMR-X SE is a fluorescent dye that can be used for labeling proteins, peptides, and other biomolecules. It has an excitation maximum at 543 nm and an emission maximum at 573 nm, making it suitable for detection with common fluorescence instrumentation.
Automatically generated - may contain errors
2 protocols using bodipy tmr x se
1
Lipid Starvation Modulates ZBTB18 Localization
2
Lipid Droplet Dynamics under Lipid Starvation
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SNB19 cells were grown in four-well chamber slides either with 10% FBS/DMEM or with 10% lipid-depleted FBS (Biowest)/DMEM for 48 h. For lipid starvation, cells were kept for an additional 2 h either in 10% lipid-depleted FBS/DMEM or in 10% FBS/DMEM. All cells were then fixed with 4% paraformaldehyde in PBS and processed for the staining. ZBTB18 was labeled with anti-ZBTB18 rabbit polyclonal primary antibody (#12714-1-AP; Proteintech) and anti-rabbit IgG (H+L) Alexa Fluor 647 (goat; Thermo Fisher Scientific) secondary antibody. Lipid droplets were stained with 0.5 μg/ml Bodipy TMR-X SE (Thermo Fisher Scientific) in 150 mM NaCl for 10 min at room temperature. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich). Pictures were acquired using a FSL confocal microscope (Olympus).
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