Fig. 6. NINJ1 is activated by cell swelling.

A–C Wild-type MEFs were stimulated with ML162 (A), MNNG (B) or H₂O₂ (C) in the presence or absence of PEG400 or PEG4000. D, F V5-NINJ1 reconstituted Ninj1 KO MEFs were treated with doxycycline overnight and the next day exposed to a 76 mOsm hypotonic shock. E Wild-type and Ninj1 KO MEFs were exposed to a 76 mOsm hypotonic shock. G, I Wild-type MEFs were stimulated with erastin (G) or FINO2 (I) in the presence or absence of PEG400 or PEG4000. H, J V5-NINJ1 reconstituted Ninj1 KO MEFs were treated with doxycycline overnight and stimulated with erastin (H) or FINO2 (J) on the next day. Plasma membrane permeabilization in function of time was measured by Sytox Green positivity (A–E, G). NINJ1 oligomerization was determined by immunoblotting after BS³ crosslinking and is representative of at least two independent experiments (D, H, J). Data in the graphs are presented as mean ± SEM of independent experiments (n = 3). Statistical significance was determined by two-way ANOVA. *P ≤ 0.05, **P ≤ 0.01, ****P ≤ 0.0001.