Fig. 7.

Staining of THP-1 cells pre-stimulated with LPS. THP-1 cells were either native (open symbols) or pre-stimulated with LPS (filled symbols), and then co-cultured with various concentrations of ABAs. a Cell surface expression of calreticulin, HMGB1 and IL-1β. THP-1 cells co-cultured with Adju-Phos, stained with antibodies and analysed by flow cytometry, collecting cell events in the P2 gate as shown in Fig. 6. Cells stained with APC-labelled anti-calreticulin (black square, white square), anti-HMGB1 (black diamond, white diamond), anti-IL-1β (black triangle, white triangle) or an isotype control (black circle, white circle). Data presented as averages of four independent experiments. b Presence of calreticulin and IL-1β on extracellular Adju-Phos particles. THP-1 cells pre-stimulated with LPS were co-cultured with various concentrations of Adju-Phos, stained with antibodies and analysed by flow cytometry, collecting extracellular adjuvant particles in the P3 gate as shown in Fig. 6. Co-culture stained with APC-labelled anti-calreticulin (black square), anti-IL-1β (black triangle) or an isotype control (black circle). Data presented as averages of four independent experiments. c Presence of calreticulin and IL-1β on extracellular Alhydrogel particles. THP-1 cells pre-stimulated with LPS were co-cultured with various concentrations of Alhydrogel, stained with antibodies and analysed by flow cytometry, collecting extracellular adjuvant particles in the P3 gate as shown in Fig. 6. Co-culture stained with APC-labelled anti-calreticulin (black square), anti-IL-1β (black triangle) or an isotype control (black circle). Data presented as averages of four independent experiments