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. 2020 Mar 16;295(18):6236–6248. doi: 10.1074/jbc.RA119.011495

Figure 3.

Figure 3.

Activation of either MyD88-dependent or TRIF-dependent signaling pathways increases MyD88-S production. A, simplified schematic depicting TLR signaling pathways. LPS is sensed by TLR4, which sequentially uses two adaptor proteins, MyD88 and TRIF, to transduce downstream signals. PAM3CSK4 stimulates TLR2, which uses the MyD88 signaling adaptor to transduce downstream signals. Poly(I:C) stimulates TLR3, which uses the TRIF signaling adaptor to transduce downstream signals. RAW264.7 cells were stimulated for 48 h with either 200 ng/ml LPS, 200 ng/ml PAM3CSK4 (P3C), 10 μg/ml poly(I:C) (pIC), or no agonist, as indicated, and either TNFα production was monitored by ELISA (B) or MyD88-L and MyD88-S expression was monitored by qPCR in conjunction with isoform-specific primers (C and D). In a separate study, RAW264.7 cells were stimulated with the same concentrations of the indicated TLR agonists, and qPCR was used to monitor MyD88-L and MyD88-S production 1 h after challenge (E and F). All experiments represent a minimum of three independent biological replicates. Data are normalized so that MyD88-L or MyD88-S expression in the absence of LPS is set to 1. *, p < 0.05.