Journal: Scientific Reports

Article Title: Aluminum adjuvants differentially induce IL-1β release in vitro yet share NLRP3 inflammasome-independent adjuvant effects in vivo

doi: 10.1038/s41598-025-34660-6

Figure Lengend Snippet: Aluminum adjuvants induce the release of inflammasome components in wild-type and Nlrp3 -/- cells. Western blot analysis of cell lysates (Lys) and supernatants (Sup) from BMDMs ( a ) and BMDCs ( b ) isolated from WT and Nlrp3 -/- mice. Cells were untreated (control), primed with LPS alone, or primed with LPS followed by stimulation for 24 h with AH or AP. Additional groups included unprimed cells stimulated with either AH or AP alone. Blots were probed for mature IL-1β (~ 17 kDa), pro-IL-1β (~ 31 kDa), caspase-1 (pro-form, ~ 45 kDa; cleaved form, ~ 20 kDa), and ASC monomers (~ 22 kDa). Ponceau S staining was used to confirm equal protein loading prior to antibody probing. Data shown are representative of two independent experiments.

Article Snippet: After blocking, membranes were incubated overnight at 4 °C with the following primary antibodies diluted in TBST (10 mM Tris-buffered saline with 0.05% Tween® 20) containing 5% bovine serum albumin (BSA): Biotinylated anti-mouse IL-1β/IL-1F2 (R&D Systems, Minneapolis, MN, cat. #BAF401; 1:1000), rabbit monoclonal anti-mouse ASC/TMS1 (D2W8U) (Cell Signaling Technology, Danvers, MA, cat. #67,824; 1:1000), mouse monoclonal anti-mouse caspase-1 (clone Casper-1, Adipogen, San Diego, CA, cat. #AG-20B-0042-C100; 1:1000), or anti- β-actin (Cell Signaling Technology, cat.#58169T).

Techniques: Western Blot, Isolation, Control, Staining