Figure EV4. MARCH5 is essential for activating the NLRP3 inflammasome.

• A
  THP‐1 cells transfected with siControl or siMARCH5 were untreated or treated with LPS alone for 4 h, ATP alone for 45 min, or LPS and ATP together.
• B
  THP‐1 cells transfected with siControl or siMARCH5 were treated with ATP for 30 min and then subjected to immunoprecipitation with ASC antibody. The levels of the indicated proteins were assessed by western blotting.
• C
  THP‐1 cells transfected with siControl or siMARCH5 were primed with LPS for 4 h and treated with 15 μM nigericin for 30 min. Triton X‐100 insoluble pellets were cross‐linked with DSS and immunoblotted with the indicated antibodies to assess ASC oligomerization.
• D
  Fluorescence microscopy images of ASC specks in LPS‐primed March5^fl/fl and March5 ^{fl/fl;Lyz‐Cre} BMDMs treated with nigericin (15 μM) for 15–20 min. ASC, green; Mitochondria, red; Nuclei, blue. White arrows indicate ASC specks. Bars, 10 μm. Independent experiments were repeated at least three times.

Source data are available online for this figure.