Skip to main content
. 2023 Sep 20;14:5832. doi: 10.1038/s41467-023-41391-7

Fig. 3. PAO1 mediated IL-1β secretion by neutrophils is NLRP3 dependent in the presence of ExoS ADPRT.

Fig. 3

Bone marrow derived macrophages (A, B) and bone marrow neutrophils (C, D) from C57BL/6 and Nlrp3-/- mice were incubated with PAO1, ∆pscD or ∆exoST. IL-1β secretion and LDH release were quantified. E GSDMD and IL-1β cleavage was examined by western blot in macrophages and neutrophils from Nlrp3-/- mice. F GSDMD and IL-1β cleavage in C57BL/6 neutrophils infected with PAO1 or mutants in the presence of the NLRP3 inhibitor MCC950. Protein cleavage was examined in lysates combined with TCA precipitated supernatants. G GTPase (GAP) and ADP ribosyltransferase (ADPRT) regions of ExoS and ExoT showing amino acids required for enzymatic function and which are also the sites of point mutations. H, I Neutrophils from C57BL/6 mice infected with PAO1, ∆exoST or mutants with indicated point mutations (ExoS, ExoT A-, G-) in the presence of MCC950. Strains expressing ExoS ADPRT are indicated. Western blots are representative of 3 repeat experiments. For panels A–D, H, I, each data point represents one independent experiments (3-6 biological repeats). Error bars are mean +/- SD. Statistical significance was assessed by 2-way ANOVA followed by Tukey’s post-test.