Fig. 4.

A RT-PCR with primers 97S and 880A on RNA extracted from HeLa cells transfected in triplicates with pC97ELsLuc and empty pUC plasmid (−) or pYTHDC1 plasmid. B Densitometric quantitation of gel image in (A). The percentage of each indicated cDNA isoform of all cDNAs in a lane in the absence of YTHDC1 (−) or in the presence of YTHDC1 overexpression is displayed. C Schematic representation of the HPV16 E6- and E7-encoding reporter plasmid pX856F. The human cytolomegalovirus promoter (CMV), the HPV16 E6 and E7 open reading frames, the HPV16 polyadenylation signal (pAL), and splice sites (226, 409, 526, and 742) are indicated. Alternatively spliced mRNAs produced by pX856F are displayed. Arrows represent RT-PCR primers 97S and x556a. D RT-PCR with primers 97S and x556a on RNA extracted from HeLa cells transfected in triplicates with pX856F and empty pUC plasmid (−) or pYTHDC1 plasmid. E Densitometric quantitation of gel image in (D). F CLIP assay was performed on cell extracts from HeLa cells transfected with pC97ELsLuc and empty pUC plasmid (−) or pYTHDC1 that produces flag-tagged YTHDC1. Immunoprecipitation of RNA–protein complexes was performed with anti-YTHDC1 antibody (a-YTHDC1) or with anti-flag antibody (a-flag). RT-PCR was performed with HPV16 primers 773S and E42as. For location of RT-PCR primers see Fig. 3C