Fig. 1: N-GSDMD is essential for nigericin-induced pyroptosis. | Cell Death & Disease

Fig. 1: N-GSDMD is essential for nigericin-induced pyroptosis.

From: ALIX mediates reversible gasdermin-D pore formation via the endosomal pathway to limit pyroptosis by active membrane repair

Fig. 1

A Immunoblot GSDMD of control and shRNA-GSDMD#1, 2, and 3 knockdown THP-1 cells. Actin was used as internal loading control. B Flow cytometry analysis of THP-1 control and shRNA-GSDMD cells at 1 h after treatment with 10 µM nigericin. The y-axis shows the percentage of cells with increased PI fluorescence intensity (PI uptake). n = 3. Significance was tested with one-way ANOVA, p** ≤ 0.01; p**** ≤ 0.0001. C Immunoblot of full length GSDMD and cleaved N-terminal GSDMD of cytoplasmic and membrane fraction of THP-1 cells after treatment with 20 µM nigericin at the indicated time points. Na + /k+ ATPase was used as marker of the membrane fraction. GAPDH was used as internal loading control. D Representative confocal microscopy images of THP-1 cells 10 and 30 min after 20 µM nigericin treatment. DAPI = Nucleus, Membrane = Biotracker 655 cytoplasmic membrane stain, Alexa 488 = cleaved N-GSDMD. The images show the maximal projection. Scale bar = 10 μM. Intensity values show the Alexa 488 channel.

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