Fig. 5. Loss of Lyn increases the susceptibility of mice to sublethal LPS-induced septic shock via a NLRP3-dependent manner.

(A) Generation of Lyn^–/– mice by CRISPR Cas9 technology. Lyn^–/– mice were generated by CRISPR-Cas9-mediated targeting of exon11 of mouse Lyn. The DNA sequence for specific gRNA is shown in red, and the PAM sequence in blue (top panel). Arrow indicates the Cas9 cleavage site. Amino acids are shown below the respective codes. The sequence of a targeted allele with 11-nucleotide deletion is presented (lower panel). The frameshift and an immediate premature stop codon TGA [X] are shown. The Lyn homozygous knockout mice with the 11bp deletion (delCTAACGTCCTG) were used in this study. (B) Splenocytes from two WT and two Lyn^–/– mice were lysed and blotted with antibodies to Lyn and actin. (C) Kaplan-Meier Survival curve of WT and Lyn^–/– mice injected with a sub-lethal dose of LPS (5 mg/kg) in the presence or absence of MCC950 (50 mg/kg) pretreatment. N = 5 each. ***P< 0.001 (Lyn^–/– + LPS vs WT + LPS or Lyn^–/– + LPS/MCC950), by log rank test. (D to F) ELISA of serum IL-1β, TNF-α, and IL-6 levels from WT and Lyn^–/– mice injected with LPS (1 mg/kg) with or without MCC950 pretreatment. N = 5. *P< 0.05（WT + LPS vs Lyn^–/– + LPS; Lyn^–/– + LPS vs Lyn^–/– + LPS/MC950, by student’s t-test. In (B to F), data are representative of three independent experiments.