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• MCF-7 cells stably expressing FADS1 and FADS2 were generated using the pcDNA3.1 expression vector system.

• Empty vector cells were used as control.

• The stable cells were grown in a specific media with 5% CO2 at 37°C.

• Cells were seeded in 60 x 15 mm cell culture dishes and grown for 48 hours.

• Cells were then incubated with 50 μM bovine serum albumin (BSA)-bound fatty acids in serum-free media.

• After 24 hours of incubation, cell pellets were harvested for fatty acid analysis.





• MCF-7 cells stably expressing FADS1 and FADS2 were generated using the pcDNA3.1 expression vector system.



• Empty vector cells were used as control.



• The stable cells were grown in a specific media with 5% CO2 at 37°C.



• Cells were seeded in 60 x 15 mm cell culture dishes and grown for 48 hours.



• Cells were then incubated with 50 μM bovine serum albumin (BSA)-bound fatty acids in serum-free media.



• After 24 hours of incubation, cell pellets were harvested for fatty acid analysis. 






• Fatty acid methyl esters (FAME) were extracted and derivatized from the harvested cell pellets.



• FAME were structurally identified using gas chromatography-covalent adduct chemical ionization mass spectrometry (GC-CACI-MS/MS).



• A specific temperature program was used for the GC.



• The MS parameters were also specifically set, including ion source temperature, interface temperature, solvent cut time, and detector voltage.



• FAME were quantified by GC-flame ionization detector (GC-FID).



• The percentage conversion of substrate to product was calculated.



• Fatty acid methyl esters (FAME) were extracted and derivatized from the harvested cell pellets.

• FAME were structurally identified using gas chromatography-covalent adduct chemical ionization mass spectrometry (GC-CACI-MS/MS).

• A specific temperature program was used for 

the GC.

• The MS parameters were also specifically set, including ion source temperature,

interface temperature, solvent cut time, and detector voltage.

• FAME were quantified by GC-flame ionization detector (GC-FID).

• The percentage conversion of substrate to product was calculated.