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. 2009 Jun 29;284(35):23405–23414. doi: 10.1074/jbc.M109.026088

FIGURE 5.

FIGURE 5.

The effect of the inducible pRb S788A on the level of stem cell marker genes and differentiation-related genes. The inducible pRb WT or pRb S788A mutant clones generated from Cdk2ap1−/− mESCs were grown in the presence or absence of LIF and with or without the induction of gene activation. The effect of the induction of pRb WT or S788A mutant expression on stem cell marker gene or differentiation-related genes was determined by real time quantitative reverse transcription-PCR analysis. A, the level of Oct3/4 and Nanog was determined in biological duplicates. The expression of pRb WT did not show any changes at the level of Oct3/4 in Cdk2ap1−/− mESCs during differentiation, but the expression of pRb S788A showed significant down-regulation of Oct3/4 regardless of LIF treatment (Oct3/4; p < 0.04). The analysis on the level of Nanog showed LIF withdrawal-dependent down-regulation only in the pRb S788A mutant (Nanog; p < 0.01). B, the levels of differentiation-related gene (Hand1, Gata6, Cdx2, and Brachyury) were compared in the inducible pRb WT and pRb S788A mESCs. The expression of Hand1 (p < 0.02) and Brachyury (p < 0.05) was most noticeably affected by the induction of pRb S788A expression, independent of LIF treatment. Statistical p values were determined by Student's t test by comparing values against the samples treated with doxycyclin and leukemia inhibitory factor (+Dox+LIF). Error bar, S.E.