Figure 3

KSRP is required for rapid degradation of a set of unstabletranscripts. (A) Immunoblot analysis of total extracts from either mock-αT3-1 (empty pSUPER-Puro vector-transfected) or αT3-1-shKSRP (pSUPER-Puro-shKSRP-transfected) cells using affinity-purified anti-KSRP and anti-α-tubulin antibodies. (B) Expression of a set of KSRP-interacting mRNAs and β2-MG (as a control), monitored by RT-PCR, in either mock-αT3-1, or αT3-1-shKSRP cells. (C) Semi quantitative RT-PCR analysis of both labile KSRP-associated mRNAs and β2-MG mRNA in either mock-αT3-1 (black lines), or αT3-1-shKSRP cells (red lines). Total RNA was isolated at the indicated times after addition of Actinomycin D. The amount of each transcript was quantitated by densitometry and plotted using a linear regression program. The values shown are averages (± SEM) of three independent experiments performed in duplicates. A quantitation of the transcripts' t(1/2) is presented in Additional file 6.