Fig. 2.

Monitoring of Texas Red-labeled dextran 10 kDa (TR10) influx in L929sAhFas iGSDME cells during apoptosis-driven secondary necrosis. a Principle of Texas Red-labeled dextran staining of L929sAhFas iGSDME cells. b–f Flow cytometry analysis of L929sAhFas iGSDME cells with (L929sAhFas iGSDME+) and without (L929sAhFas iGSDME-) doxycycline-induced GSDME expression during apoptosis-driven secondary necrosis. b Representative plots of L929sAhFas iGSDME cells untreated and after treatment with anti-Fas for 8 h. c Levels of Texas Red single-positive cells (TR10+/SB−) in L929sAhFas iGSDME cells upon anti-Fas treatment. d Levels of Texas Red and SB double-positive (TR10+/SB+) cells in L929sAhFas iGSDME cells upon anti-Fas treatment. e Fraction of Texas Red positive (TR10+) and Texas Red negative (TR10−) cells in the SB− population. f Fraction of Texas Red positive (TR10+) and Texas Red negative (TR10−) cells in SB+ population. Dox doxycycline; GSDME gasdermin E; LsFas L929sAhFas; SB SYTOX Blue; NTC non-treatment control; TR Texas Red