(A) Western blot analysis of IL-1β and caspase-11 protein expression in
BMDMs following treatment with LPS (500 ng/ml), poly (I:C) (5 μg/ml),
Pam3CSK4 (1 μg/ml), or IFN-γ (10/ng/ml) for six hours. (B) Q-PCR
analysis of IL-1β and caspase-11 mRNA expression in BMDMs following
treatment with LPS (500 ng/ml), poly (I:C) (5 μg/ml), Pam3CSK4 (1
μg/ml), or IFN-γ (10/ng/ml) for six hours. (C) Western blot
analysis of indicated proteins in BMDMs recovered from mice with the indicated
genotypes following LPS electroporation or E. coli
(MOI=25) infection for 16 hours. Sup= supernatants. (D) In
parallel, cytotoxicity (LDH release) and levels of IL-1β and HMGB1 in
the supernatant were assayed (n=3 wells/group; data expressed as means
± SD, *, P<0.05, ANOVA LSD
test). (E) Western blot analysis of LPS (500 ng/ml)-primed indicated BMDMs
following treatment with ATP (5 mM) for one hour. Sup=supernatants. (F)
In parallel, cytotoxicity and levels of IL-1β and HMGB1 in the
supernatant were assayed (n=3 wells/group; data expressed as means
± SD, *, P<0.05, ANOVA LSD
test).