MYC activation and Rb deletion drive HCC development in the liver of adult mice.

<p><b>A.</b> Strategy to produce <i>MYC</i> and <i>MYC/Rb</i> mutant tumors using <i>Rb^lox/lox</i> and TRE-<i>MYC</i> LAP-<i>tTA</i> mice. MYC is activated in the liver by the removal of doxycycline from the drinking water while <i>Rb</i> is specifically deleted in the liver by splenic injection of Ad-Cre. Ad-GFP is used as a control. <b>B.</b> RT-qPCR analysis of <i>MYC</i> RNA levels in control livers (<i>Rb^lox/lox MYC^Off</i>, n = 2) and in <i>MYC</i> and <i>MYC/Rb</i> mutant tumors (n = 3). <b>C.</b> Immunoblot analysis of MYC protein levels in control livers (<i>MYC^Off</i>) and in <i>MYC</i> and <i>MYC/Rb</i> mutant tumors (two independent tumors each, T1 and T2). Actin serves as a loading control. <b>D.</b> Genomic PCR analysis for the deleted allele of <i>Rb</i> (<i>Rb^Δ</i>) using DNA from <i>MYC</i> and <i>MYC/Rb</i> mutant tumors. <i>Actin</i> serves as a positive PCR control. <b>E.</b> RT-qPCR analysis of <i>Rb</i> RNA levels in <i>MYC</i> (black, n = 3) and <i>MYC/Rb</i> (grey, n = 3) mutant tumors.