Figure 6.

Characterization of TLR7-activating rRFs

(A) Normalized read counts for the three selected rRFs. (B) After DOTAP-mediated endosomal delivery of the indicated RNAs, cytokine mRNAs were quantified using RT-qPCR. The graph displays average values of relative abundance, with the data for ssRNA41 set as 1. Error bars indicate the SD. (C) After DOTAP-mediated endosomal delivery of the indicated RNAs, the culture medium was analyzed using ELISA to measure the concentrations of the indicated cytokines. Results from two independent experiments are shown as separate bars. (D) The DOTAP experiments were performed using two different TLR7 KO THP-1 cell clones (#1 and #2) followed by quantification of cytokine mRNAs using RT-qPCR. The average values of relative abundance are shown, with data for ssRNA41 set as 1. Error bars indicate the SD. (E) The indicated wild-type and mutant rRFs were subjected to DOTAP experiments, followed by quantification of TNF-α mRNA levels. (F and G) After DOTAP-mediated endosomal delivery of the indicated RNAs, HMDMs were subjected to bacterial infection and invasion assay. Representative images of plates with E. coli colonies (F) and bar graphs of the counted colony numbers (G) are shown. Results from two independent experiments are shown as separate bars.