Fig. 3: Nanobodies preventing oligomerization still allow membrane localization of overexpressed GSDMDNT. | Nature Communications

Fig. 3: Nanobodies preventing oligomerization still allow membrane localization of overexpressed GSDMDNT.

From: Antagonistic nanobodies implicate mechanism of GSDMD pore formation and potential therapeutic application

Fig. 3

A HEK293T cells stably expressing the indicated VHH-EGFP fusions were transfected with expression vectors for GSDMD-mCherry I104N (left) or GSDMDNT-mCherry I104N (right) and analyzed by live-cell confocal imaging. Data representative of three independent experiments are shown. Yellow arrows indicate GSDMDNT-mCherry in intracellular vesicular structures. B HEK293T cells stably expressing VHHGSDMD-1-EGFP were transfected with expression vectors for the plasma membrane marker emiRFP670-CAAX as well as the indicated GSDMD variants fused to mCherry. Representative images are shown on the left. The distribution of the indicated GSDMD variants in cells positive for emiRFP670 and mCherry were enumerated and average values from n = 3 independent experiments with at least n = 30 cells per condition are displayed  ±  SEM on the right. PM, plasma membrane. Scale bars, 10 µm.

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