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. 2012 Feb 13;209(2):225–233. doi: 10.1084/jem.20111044

Figure 1.

Figure 1.

Native E. coli tRNATyr lacks immunostimulation in PBMCs. (A) PBMCs were stimulated with E. coli (E.c.) tRNAs, tRNAPhe from eukaryotic S. cerevisiae (S.c.), mammalian (mam.) tRNAPhe, or mammalian tRNALys3 and analyzed for secretion of IFN-α. Data are normalized to 1 µM CpG2216 (n = 3–10; mean + SEM). (B) PBMCs or PBMCs depleted of pDCs were stimulated with E. coli tRNAPhe and analyzed as in A (mean of triplicate values + SD). (C) PBMCs were preincubated with 1 or 5 µM chloroquine (Chl) and stimulated with 1 µg/ml IVT of E. coli tRNAPhe. (D) PBMCs were stimulated with 300 ng/ml E. coli tRNALys in the absence or presence of a 1:1 or 1:2 ratio of the TLR7-blocking RNA oligonucleotide 2mA. (C and D) One of two experiments is shown (mean of duplicate values + SD). (E) PBMCS were treated with siRNA against TLR7 and subsequently stimulated with 1 µg/ml E. coli tRNALys or 1 µg/ml R848 and tested for IFN-α secretion (one of two experiments is shown; mean + SD). (F) Huh7.5 cells harboring an IRF3 reporter gene were transfected with either RIG-I or MDA5. Cells were stimulated with 1 µg/ml E. coli tRNALys; 0.2 µg/ml of 400-bp ds IVT RNA or poly (dI:dC) was used as control, and reporter gene activity was measured (n = 2; mean + SD). RLU, raw light units.