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. 2018 Mar 27;46(10):5029–5049. doi: 10.1093/nar/gky227

Figure 9.

Figure 9.

(A) sLuc activity induced from pBELsLuc in HeLa cells co-transfected with pBELsLuc and the indicated plasmids. (B) 3′-RACE assay on total RNA extracted from HeLa cells transfected with pBELsLuc reporter plasmid and empty vector or a plasmid expressing hnRNP C1. The primers specifically detect mRNAs polyadenylated at HPV16 pAE or HPV16 pAL. The pAE- or pAL-3′-RACE products were quantified and the levels detected in cells transfected with hnRNP C plasmid were divided by the levels detected in cells transfected with empty plasmid (vector). The ratios are indicated below the gels. (C) RT-PCR on HPV16 E4 mRNAs or L1 and L1i mRNAs (D) produced from pBELsLuc transiently transfected into HeLa cells in the presence of empty vector or plasmid expressing hnRNP C1. The RT-PCR products were quantified and the levels detected in cells transfected with hnRNP C plasmid were divided by the levels detected in cells transfected with empty plasmid (vector). In case of the L1 mRNAs, the ratios between the L1 and L1i splice variants of the L1 mRNAs in cells transfected with hnRNP C plasmid or empty plasmid (vector) are shown. The ratios are indicated below the gels. GAP, GAPDH mRNAs. (E) Schematic representation of genomic HPV16 plasmid pHPV16ANSL (32,59). LoxP sites and HPV16 early (p97) and late (p670) promoters and early (pAE) and late (pAL) poly(A) signals are indicated. The effect of the cre recombinase on these plasmids is illustrated (59). (F) sLuc activity produced in HeLa cells transfected with pHPV16ANSL in the absence or presence of various concentrations of hnRNP C-expression plasmid. (G) 3′-RACE assay on total RNA extracted from HeLa cells transfected with pHPV16ANSL and empty vector or hnRNP C1- or Fip1- expression plasmid. The 3′-RACE primers specifically detect mRNAs polyadenylated at HPV16 pAE or HPV16 pAL. The pAE- or pAL-3′-RACE products were quantified and the levels detected in cells transfected with hnRNP C plasmid or Fip1 plasmid were divided by the levels detected in cells transfected with empty plasmid (vector). The ratios are indicated below the gels.