CD40L protein expression and IL-12 cytokine secretion in DC cultures transfected with various CD40L RNAs. Panel A: intracellular staining of DCs with anti-CD40L antibody at 4 hours post-electroporation with the RNAs. Panel B: cytokine levels measures in DC culture supernatants after overnight incubation. GFP is used as a negative control RNA. Type of cap analogue is indicated by shading back: ARCA and hatched m7G. All RNAs in this experiment were capped co-transcriptionally. All RNAs were polyadenylated in a post-transcriptional reaction and contained >150 nucleotide long poly(A) tail, except for CD40L+5'UTR 64A. For this RNA poly(A) tail was obtained in a transcription reaction by transcribing from templates containing poly(T) stretch after CD40L coding region. 5'UTR 64A+polA tail designates the RNA which in addition to 64 long poly(A) tail were post-transcriptionally polyadenylated. In this case poly(A) tail was >210 nucleotide long.