Extended Data Fig. 4: Sequence differences in GSDMB isoforms and characterizations of their functions.
From: Structural mechanisms for regulation of GSDMB pore-forming activity
a, Sequence alignment of GSDMB isoforms. Domain annotation is shown on top of the sequences. The exon 6 and 7 regions are labeled and highlighted by black boxes. GZMA cleavage site Lys244 (numbered after GSDMBiso3) is highlighted in blue background. The site for inserting a PPase-cleavage sequence is marked by a green arrow. Identical residues are in red background. Numbers of starting residues are indicated on the left. b, Gel-filtration chromatography analyses of complex formation between IpaH7.8 and different GSDMB isoforms, along with SDS-PAGE of the complex elution fractions. c, Assay of GSDMBIso3 in IpaH7.8 blocking of GZMA-induced pyroptosis. Flag-IpaH7.8 (WT or C357S) was transfected into 293T cells stably expressing GSDMBIso3; GZMA was electroporated into the cells. LDH release-based cell death data are means (bars) of three replicates (circles). Cell lysates were subjected to immunoblotting. d, 293T cells expressing different GSDMBPP isoforms were electroporated with PPase to cleave the GSDMBPP. Morphological examination of cell pyroptosis is shown. Scale bar, 25 μm. Data (b−d) are representative of three independent experiments. See Supplementary Fig. 1 for gel source data.
