Of note, arrest in mitosis with 2ME2 induced only a slight reduction in Smad3 mRNA, which failed to reach statistical significance (measured by qRT-PCR, 0.8±0.21 fold of untreated, n = 6; p>0.2, 1-tailed t-test), suggesting a minimal contribution of a reduction in transcription to the observed decrease of tSmad3 levels.
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Differential regulation of Smad3 and of the type II transforming growth factor-β receptor in mitosis: implications for signaling.
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Proteasome inhibition (3 h) resulted in a marked accumulation of pSmad3C (∼2.5 fold), a lesser increase in tSmad3 (which did not reach statistical significance), and no increase in pSmad3(179) ( Figure 2E ).