Table 1. Pyruvate carboxylase knockdown lines show decreased incorporation of glutamine carbon into anaplerotic products and into lipid.

Cells were maintained for 24 h in RPMI cell culture medium containing 5 mM glucose and without pyruvate and then harvested and incubated in suspension for 45 min in the presence of 10 mM BCH and 10 mM [U-¹⁴C]glutamine (specific radioactivity 2.5 mCi/mmol) followed by extraction of the radioactivity incorporated into anaplerotic products and a lipid fraction. Results are expressed as the mean ± SE from four replicate incubations.

Cell Line	Glutamine Incorporation
	(nmol/mg cell protein/h
	Anaplerotic Products	Lipid Fraction
U6 (Control)	0.64 ± 0.05	0.20 ± 0.01
PC118/1973	0.49 ± 0.03a	0.13 ± 0.01b
PC118/3064	0.34 ± 0.015b	0.11 ± 0.002c

^ap < 0.05,

^bp < 0.005 and

^cp < 0.001 vs control