Extended Data Fig. 9: NINJ1 does not affect full PMR under mechanical strain in LLO induced cell death.
From: NINJ1 regulates plasma membrane fragility under mechanical strain
a, NINJ1 KO and parental THP-1 cells were treated with 500 ng ml−1 LLO and stained with CellBrite (red), DRAQ7 (yellow) and Hoechst (blue) at 1 h, 2 h, 8 h, 12 h and 24 h later. b, Quantification of LDH release, percentage of DRAQ7+ cells and DRAQ7+ cells with bare nuclei at various time points after LLO treatment. n = 3 for each group. Data were analysed using 2-way ANOVA followed by Bonferroni corrections. ** p < 0.01 versus parental+LLO. c, NINJ1 KO and parental THP-1 cells were treated with LLO for 1 h before flow stimulation at the shear rate 829 s−1. Cells were stained after 30 min as previously described. d, Cells were treated with 500 ng ml−1 LLO for 1 h, then subjected to flow at 829 s−1 in shear rate. Measurement of the LDH release, percentage of fully ruptured parental and KO cells were carried after 30 min. Data are from 3 trials. To quantify DRAQ7 staining and ballooned cell percentage, 250 ~ 300 cells examined for each trial, and analysed using two-sided unpaired Student’s t-test, n.s., not significant versus parental. All data are presented as mean ± s.e.m. Scale bars, 25 µm.
