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. Author manuscript; available in PMC: 2020 Oct 9.
Published in final edited form as: Sci Signal. 2019 Oct 8;12(602):eaao3829. doi: 10.1126/scisignal.aao3829

Fig. 5. Regulation of TLR-triggered signaling and cytokine production by Lyn requires CD11b and Src-family kinases Hck and Fgr.

Fig. 5.

(A to C) BMDCs or BMMs from the indicated mice were stimulated with 100 ng/ml Pam3CSK4 for the indicated time. Cells were lysed and immunoblotted with the indicated antibodies. (D) BMDCs from the indicated mice were either centrifuged onto the bottom of tissue culture treated plates (att) or left in suspension (unatt) with or without the addition of antibody to CD11b (Mac-1) for 30 mins (αCD11b). Cells were stimulated with 100 ng/ml of Pam3CSK4 for the indicated times and supernatants were analyzed for TNF-α by ELISA. (E and F) BMDCs from the indicated mice were stimulated with the indicated amount of Pam3CSK4 and LPS for 3 hours. Cell supernatants were analyzed for TNF-α by ELISA. (G) BMDCs from the indicated mice were left untreated or stimulated with 100 ng/ml Pam3CSK4, 100 ng/ml LPS for 30 mins. Cells were lysed and immunoprecipitated with CARD9 antibody, followed by immunoblotting with phospho-threonine (P-TxR) antibody or CARD9 antibody. Densitometry analysis of the western blots are shown in fig. S1. Blots are representative of at least 3 independent experiments with cells pooled from 3 to 5 mice per group. (H) DSS-induced weight loss was monitored and analyzed as in Fig 1A. * p<0.05, one way ANOVA with Tukey HSD test for (D-F) or Student’s t-test (H).