Extended Data Fig. 8: NINJ1 facilitates full PMR under mechanical strain in intracellular LPS induced cell death.
From: NINJ1 regulates plasma membrane fragility under mechanical strain
a, NINJ1 KO and parental THP-1 cells were electroporated with LPS and stained with CellBrite (red), DRAQ7 (yellow) and Hoechst (blue) at 1 h, 8 h, 16 h and 24 h after LPS treatment. Almost all cells showed DRAQ7 staining, indicating large breaches on the plasma membrane, yet still displayed intact ballooned shape without full rupture. b, Quantification of LDH release, percentage of DRAQ7+ cells and percentage of ballooned cells at various time points after electroporation of LPS. n = 3 for each group. c, NINJ1 KO and parental THP-1 cells were electroporated with LPS 1 h before being subjected to flow treatment at shear rates ranging from 829 s−1 to 2073 s−1. A significantly lower percentage of NINJ1 KO cells show DRAQ7+/Hoechst+ bare nuclei, indicative of full PMR, after being subjected to flow. In all assays, data are from 3 trials. To quantify DRAQ7 staining and ballooned cell percentage, 250 ~ 300 cells examined for each trial. Data were analysed using 2-way ANOVA followed by Bonferroni corrections and presented as mean ± s.e.m. Scale bars, 25 µm.
