Fig 6. The cell death defect in Card19lxcn BMDMs is independent from cytokine release.
(A-E) B6 (black bars) and Card19lxcn (red bars) BMDMs were primed with LPS for 3 hours and treated with extracellular ATP, infected with S. Tm, Yptb, or the Yptb ΔEJK strain lacking effectors YopE, YopJ and YopK. Supernatants were harvested 2 hours post-infection or ATP treatment and analyzed by ELISA for release of (A) IL-1α, (B) IL-1B, (C) IL-6, (D) IL-12, and (E) TNF-α. Mean ± SEM is displayed. Each figure is representative of 3 or more independent experiments. (F-H) B6 BMDMs were primed with LPS for 4 hours and infected with ΔoatA S. aureus or S. Tm. (F) Lysates were harvested at indicated times, run on SDS-PAGE page and analyzed by western blotting for Caspase 1, GSDMD, CARD19, or Actin (loading control). Supernatants were harvested at indicated time points and analyzed by (G) LDH for cytotoxicity and (H) ELISA for release of IL-1β. Each figure is representative of two independent experiments.