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. 2023 Jan 26;14:1113883. doi: 10.3389/fimmu.2023.1113883

Figure 4.

Figure 4

Hepatic inflammatory monocyte (IM) and monocyte-derived macrophage (MDM) caspase-11 deficiency protective against pyroptosis, WT bone marrow transplantation to Casp11–/– mice restored IM and MDM pyroptosis. 8-10 weeks old male WT and Casp11–/– mice were fed HFD for 12 weeks. (A) Western blot for noncanonical pyroptosis mediators. (B) Liver IL-1β concentrations. (C) Experimental design for bone marrow transplantation (BMT). (D) Representative flow cytometry gating of hepatic macrophages (HMΦ, Green, CD45+ > CD11b+ Ly6G- > Ly6Clow MHCIIhigh) inflammatory monocytes (IM, Red, CD45+ > CD11b+ Ly6G- > Ly6Chigh MHCIIlow). (E) Gating strategy for designing pyroptosis populations. HMΦs and IMs gated on GSDMD MFI vs. Casp11-Inhibitor MFI. RED: GSDMD MFI for IM (CD45+ > CD11b+ Ly6G- > Ly6Chigh MHCIIlow). GREEN: Percentage of the parent for HMΦ pyroptosis gating CD45+ > CD11b+ Ly6G- > Ly6Chigh MHCIIlow > Casp11 Activity vs GSDMD). (F) Schematic diagram showed that WT mice had more monocyte migration and more hepatic pyroptosis, however, Casp11–/– had less monocyte migration and hepatic pyroptosis resulting in an unchanged total number of hepatic macrophages. Statistical Analysis: Flow cytometry data was analyzed with FlowJo, and statistical analysis was performed using Prism. One-Way ANOVA. One-Way ANOVA. *p < 0.05, **p < 0.001, ***p < 0.0001 ****p < 0.0001. ns, Non-significant.