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. 2020 Apr 27;217(7):e20191736. doi: 10.1084/jem.20191736

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© 2020 Tenthorey et al.

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Figure 2. — NLRC4 phosphorylation is neither sufficient nor strictly required for response to cytosolic flagellin. (A–E) Macrophages were either left unprimed (A and B) or primed 4 h with 1 µg/ml Pam3CSK4 (C–E) and then treated with 4 µg/ml PA and the indicated dose of LFn-FlaA to activate NLRC4. LDH release (A and C), PI uptake (B and D), or IL-1β release (E) was measured after 4 h. All data are representative of three independent experiments (three biological replicates per experiment). Mean ± SD; n.d., not detectable. Repeated measures two-way ANOVA with Dunnett’s multiple comparisons post-test, all genotypes vs. C57BL6/J (or Nlrc4^S533A/S533A vs. Nlrc4^−/− as indicated), *, P < 0.05, **, P < 0.005, ***, P < 0.0005, ****, P < 0.00001; ns, not significant.