Figure Legends:
Figure 1 . Asthma triggers cleave GSDMB. (A) Western blot
showing cleaved GSDMB from differentiated airway epithelial cell lysates
mixed with HDM extracts or 293T cell lysates (transfected with GSDMB)
mixed with increasing concentrations of (B) HDM extracts and (C) mold or
cockroach extracts. (D) Cleavage of recombinant GSDMB by HDM allergen.
Box marks the fragment subjected to sequencing. (E) Upper panel: Results
of the Edman Sequencing of the first 10 amino acids. Residues that were
identified with low degree of certainty are in parentheses. Lowe panel:
Diagram depicting the position of the identified amino acids in the
GSDMB protein. (F) In vitro cleavage assay in 293T lysates
transfected with WT-GSDMB or K244A and incubated with HDM extracts.
Anti-GSDMB (A) or anti-FLAG antibody (B,C,F) was used to detect
full-length and cleaved GSDMB.
Figure 2. GSDMB-NT generated by HDM cleavage induces
pyroptosis. (A) Schematic diagrams showing the WT GSDMB and the
resulting GSDMB-NT fragments (NT-K244 and NT-K231var) after HDM
cleavage. (B) Representative images of cells transfected with indicated
plasmids. Blue arrowheads mark pyroptotic cells. (C) LDH release assay
from 293T cells transfected with the indicated plasmid constructs.
Results indicate mean +/- SEM of n=6 from a representative experiment.
*p <0.05.