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. Author manuscript; available in PMC: 2013 Sep 1.
Published in final edited form as: J Cell Physiol. 2012 Sep;227(9):3310–3316. doi: 10.1002/jcp.24026

Fig 3. Inflammasome silence blunted ceramide induced cytokine production in THP-1.

Fig 3

(A)Inflammasome activation was assessed by analyzing caspase-1 cleavage using Western blot analysis. shRNA silencing of NALP-3 abrogated ceramide-induced caspase-1 cleavage (upper panel) as well as secretion of IL-1β in ceramide treated (30 μM) THP-1 cells (lower panel, graph). ATII cells were grown in 24-well transwell plates until the cellular monolayers reached confluence. THP-1 cells were added to the appropriate ATII monolayers, cocultured for 6 h, and exposed to ceramide for 12 h. In some experiments, THP-1 cells were transfected with control shRNA or NALP-3 shRNA and then added to a monolayer of ATII cells. The media contents were collected and (B) IL-1β, (C) TNF-α and (D) IL-6 proinflammatory cytokines were analyzed in extracellular media by ELISA. Asterisks indicate significant differences compared to the AT-II (***P < 0.001) and compared to control shRNA (mean s.d.; **P < 0.01). The noted experiments are representative of a minimum of three similar evaluations.