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. 2004 Oct 11;167(1):35–41. doi: 10.1083/jcb.200406136

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Copyright © 2004, The Rockefeller University Press

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Figure 4. — Expression of enzymes that function in the CDP-choline pathway of PtdCho synthesis. Total RNA was prepared from NIH-3T3 fibroblasts harvested at 24 and 48 h after transduction with the indicated retroviral vectors. (A) The relative levels of expression of the CCTα, β2, and β3 isoform cDNAs at the 48-h interval were measured by quantitative real-time PCR. The amount of target RNA was normalized to the endogenous GAPDH reference and related to the amount of target CCTα in NIH-3T3, which was set as the calibrator at 1.0 (superscript a). The mean ± SD (superscript b) of triplicate determinations is shown. (B) Northern blot analysis using ³²P-labeled cDNA probes specific for CEPT1 and CPT1, the two genes encoding enzymes possessing CPT activity; ERdj3, a known XBP1 target as a positive control; and γ-actin as a loading control.