Fig. 3: TAGAP-binding partner EPHB2 is important for antifungal signaling activation.

a HEK293T cells were transfected with indicated plasmids, and cell lysates were immunoprecipitated with anti-Flag antibody, followed by immunoblot analysis for indicated proteins. b Flag-TAGAP stable transfected THP-1 cells were differentiated with PMA (25 ng/mL) for 3 days, followed by immunoprecipitation by Flag antibody. Protein elution was analyzed by mass spectrometry analysis. Graph represents mass spectrometry result of TAGAP-interacting proteins. c Human THP-1 cells were non-polarized or polarized by adding PMA (25 ng/mL) for 3 days, and were stimulated with heat-killed C. albicans (MOI = 2) for 0, 15, and 30 min, followed by western blot analysis of indicated proteins. d, e HEK293T cells were transfected with indicated plasmids, and cell lysates were immunoprecipitated with anti-Flag antibody, followed by immunoblot analysis for indicated proteins. f Control siRNA or EPHB2 siRNA-transfected BMDMs were stimulated with Curdlan (100 μg/ml) for 3 h, followed by real-time PCR analysis of indicated genes. g–i THP-1 cells infected with control gRNA or EPHB2-gRNA were stimulated with heat-killed C. albicans (MOI = 2), Curdlan (100 μg/ml) or α-Mannan (100 μg/ml) for the indicated times, followed by western blot analysis of indicated proteins. j THP-1 cells infected with control gRNA or EPHB2-gRNA were stimulated with heat-killed C. albicans (MOI = 2) for the indicated times, followed by real-time PCR analysis for indicated genes. P < 0.05; **P < 0.01; ***P < 0.001 based on two-sided unpaired t test f, j. Real-time PCR data of f and j were collected from two independent experiments. All error bars represent SEM of technical replicates. Data are representative of three independent experiments.