Fig. 1

Identification of the p40-EBI3 protein in mice. A Splenocytes isolated from normal BALB/c mice were stimulated with LPS, zymosan, or Con A or were untreated (Nil). The expression levels of p40-EBI3 in culture supernatants were detected using ELISA and converted into OD (405 nm) and pg/mL values. B Detection of p40-EBI3 protein expression in splenic non-T cells by immunoprecipitation with western blot analysis. Splenic non-T cells isolated from WT BALB/c mice were stimulated under different conditions for 3 days. Prepared cell lysates were immunoprecipitated with an anti-EBI3 antibody and subjected to western blotting with an anti-p40 antibody. C CD11c⁺ splenic dendritic cells isolated from normal BALB/c mice were stimulated under different conditions. The expression levels of the p40-EBI3 protein in culture supernatants were determined by ELISA. D Detection of p40-EBI3 protein expression in splenic non-T cells by EBI3 immunoprecipitation with p40 western blot analysis. Splenic non-T cells isolated from WT BALB/c mice were stimulated with 5 μg/mL ConA for 3 days. The p40-EBI3 protein was detected in the cell lysates and culture supernatant. E Schematic map of the p40-EBI3 plasmid vector (details described in the Methods section). F HEK293 cells were transfected with the p40-EBI3 plasmid vector. The expression levels of p40, EBI3, and FLAG in transfected cells were determined by western blotting. G The expression levels of p40-EBI3 in the culture supernatants in E were determined by ELISA (*p < 0.05, **p < 0.01, ***p < 0.001).