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. 2006 Jul 10;103(29):10961–10966. doi: 10.1073/pnas.0603804103

Fig. 2.

Fig. 2.

Effects of the Poc and Lkd mutations in vivo and ex vivo. (A) The Poc mutation confers sensitivity to MALP-2-induced lethal toxicity. Wild-type, Poc, and MyD88-deficient mice were injected i.p. with 3 μg of MALP-2 and 20 mg of d-galactosamine. Survival was monitored over a period of 3 days (no change was observed after 48 h), and the data are expressed as a Kaplan–Meier plot (P < 0.0001). (B and C) The Poc allele of MyD88 supports resistance to skin infection caused by S. pyogenes. Mice were each injected s.c. with S. pyogenes (5 × 105 colony-forming units). During a 5-day period, bacterial growth was monitored daily with the Xenogen IVIS imaging system. (C) Luminescence (photons emitted per second) was measured in a defined and constant region of interest. Data are shown as means ± SEM0 (n ≥ 4 mice). (D and E) Poc mice are hypersusceptible to L. monocytogenes infection. Mice were injected i.v. with L. monocytogenes as described in Materials and Methods. Bioluminescence imaging was performed by using the IVIS Imaging System. The survival of these mice was monitored during a 7-day period. P values refer to comparison with wild type (E). (F and G) Poc mice are hypersusceptible to mouse CMV infection. (F) Viral titers, expressed as log plaque-forming units per spleen, were determined in mice 5 days after i.p. inoculation with 5 × 105 plaque-forming units of mouse CMV. (G) Blood was collected 36 h after infection, and the concentrations of type I IFN in serum were analyzed by ELISA. (H) Macrophages from Poc or MyD88−/− mice were treated with either MALP-2 or Pam2CSK4 for the indicated times. Cells were lysed and analyzed by immunoblotting with antibodies indicated. (I) MyD88Poc interacts with TLR2 after MALP-2 treatment. HEK-293 cells were transiently transfected with M2-Flag-tagged TLR2 and HA-tagged WT-MyD88 or MyD88Poc and treated with Pam3CSK4 or MALP-2 for the indicated times. Cells were lysed and immunoprecipitated with M2-Flag followed by immunoblotting with antibody against the HA. The expression of transfected MyD88 and TLR2 was examined in the whole-cell extracts (WCE).