Fig. 3
NINJ1 mediates IAV-induced cell lysis without compromising cell death. a, b Representative images (a) and quantification (b) of PI+ cells in wild-type (WT) and Ninj1-/- BMDMs at 12 hpi. The arrows indicate unlysed cells exhibiting a balloon-like morphology. Scale bars, 100 μm and 40 μm in the enlarged image. c–f LDH release (c), immunoblots of the indicated proteins (d), silver staining of supernatants (e), and cell viability (f) in BMDMs (genotypes as above) at 12 hpi. g Concentrations of TNF-α in supernatants from WT and Ninj1-/- BMDMs at 16 hpi. h Concentrations of IL-1β in supernatants from WT, Ninj1-/-, and Gsdmd-/- BMDMs at 16 hpi. i qRT‒PCR analysis of IAV-NP vRNA in WT and Ninj1-/- BMDMs at 12 hpi (normalized to WT group). j Immunoblots of NINJ1 in BMDMs with/without glycine (10 mM) at 12 hpi, BS3-crosslinked. k qRT‒PCR analysis of Ninj1 expression in BMDMs with/without glycine (10 mM) at 12 hpi (normalized to uninfected controls). l LDH release in WT and Ninj1-/- BMDMs with/without glycine (10 mM) at 12 hpi. m Cell viability of BMDMs with/without glycine (10 mM) at 12 hpi. Data are representative of at least two independent experiments and are presented as mean ± SD. Student’s t-test (f, i, and m) or two-way ANOVA (b, c, g, h, k, and l) was applied. ns, not significant; ****p < 0.0001
