Time course of NMT knockdown. Time course of NMT knockdown. A. SK-OV-3 cells were treated with 10 nmol/L NMT1-1 siRNA, 10 nmol/L NMT2-4 siRNA, or 10 nmol/L each of NMT1-1 and NMT2-4 siRNA for 48 hours as described in Materials and Methods. Western blots were prepared using 50 μg total protein harvested from the cells and were first probed with either anti-NMT1 or anti-NMT2 antibodies, stripped, and then reprobed with a pan-actin antibody. The standards are Magic Mark Western standards (Invitrogen) that are visualized by anti-mouse horseradish peroxidase–conjugated secondary antibodies. B. A titration of HT-29 total cell protein (10-100 μg) probed for NMT1 and NMT2 and quantified on the Fujifilm imager. The graph shows the signal intensity of each band versus the concentration of total cell protein loaded for both NMT1 and NMT2 blots. Charles E. Ducker et al. Mol Cancer Res 2005;3:463-476 ©2005 by American Association for Cancer Research