Post-transcriptionally capped CD40L RNAs are 100% capped and induce the highest level of IL-12 secretion. Panel A. Oligonucleotide-directed RNaseH cleavage products from the CD40L MET1 capping efficiency assay analyzed by denaturing polyacrylamide gel electrophoresis and autoradiography. Cleavage products from uncapped and capped RNA molecules are denoted by arrows. The control RNA is co-transcriptional ARCA-capped ΔXE-MET1 RNA. Capping efficiency measurements are indicated at the bottom of the gel image. Panel B: IL-12 cytokine secretion induced by ΔXE-MET1 RNA transfection as measured by ELISA. The resulting cap of each RNA is indicated by colour of the bars and as indicated, the type of cap (I or 0) is indicated below the graph. CD40L WT Reference RNA prepared with an ARCA cap by co-transcriptional cap analogue incorporation. GFP is a negative control RNA. All RNAs were polyadenylated post-transcriptionally and have >150 nucleotide tails. non-standard format).