1. Quantification of NMT knockdown.
Quantification of NMT knockdown. SK-OV-3 cells were treated with 10 nmol/L NMT1-1 siRNA, 10 nmol/L NMT2-4 siRNA, 10 nmol/L each of NMT1-1 and NMT2-4 siRNA, 20 nmol/L negative control siRNA (Neg. Cont.), or no siRNA in the absence and presence of LipofectAMINE 2000 (LF2000) for 48 hours as described in Materials and Methods. Western blots were prepared using 50 μg total protein harvested from cells and were first probed with either anti-NMT1 (A) or anti-NMT2 antibodies (B), stripped, and then reprobed with a pan-actin antibody. The standards are Magic Mark Western standards that are visualized by anti-mouse horseradish peroxidase–conjugated secondary antibodies. The graphs represent the ratio of NMT1 or NMT2 to actin in each lane. Northern blots (C) were prepared using 10 μg mRNA per lane from the same cell treatments and were probed with NMT1, NMT2, and β-actin probes as described in Materials and Methods.
Charles E. Ducker et al. Mol Cancer Res 2005;3:
©2005 by American Association for Cancer Research