Inactivation of Icmt inhibits transformation by oncogenic K-Ras and B-Raf
J. Clin. Invest. Martin O. Bergo, et al. 113:539 doi:10.1172/JCI18829 [
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Figure 2Analyzing the effect of
Icmt inactivation on cell growth. (
a) Anchorage-dependent growth of
Icmtflx/flx and
IcmtΔ/Δ cells. Equal numbers of immortalized
Icmtflx/flx cells and the derivative
IcmtΔ/Δ cells (lines A and B) were plated onto 96-well plates (
n = 12 wells per cell line), and cell growth was assessed with the Cell Titer 96 AQueous One Solution Cell Proliferation Assay (Promega). (
b) Southern blots illustrating the increase in the ratio of
Icmtflx to
IcmtΔ bands during the growth of a mixed population of
Icmtflx/flx and
IcmtΔ/Δ cells. Mixed populations of
Icmtflx/flx and
IcmtΔ/Δ cells were passaged at a 1:10 ratio every 3 days. DNA was harvested and analyzed with Southern blots at the indicated passages. (
c) Southern blot of liver DNA from three
Icmtflx/flxMx1-Cre mice before and after hepatocyte proliferation, which was induced by a partial hepatectomy. The ratio of
Icmtflx to
IcmtΔ band intensity increased after liver regrowth, indicating that
Icmtflx/flx hepatocytes contributed more to liver regrowth than the
IcmtΔ/Δ hepatocytes. Quantification of data from five mice revealed that the
Icmtflx/
IcmtΔ ratio increased 201% ± 26% after liver regeneration (
P < 0.01).
