BIRB 796 and dasatinib suppress TLR-activated TNFα gene expression in primary and patient-derived FA cells. Means ± SD are shown. All P values were calculated using the Student t test. (A) BIRB 796 and dasatinib inhibit constitutive TNFα secretion in the patient-derived FANCC^−/− lymphoblast cell line HSC536N. HSC536N and HSC536N/FANCC (complemented with normal FANCC cDNA) were cultured in medium alone (control) or medium treated with BIRB 796 (500nM) or dasatinib (500nM) for 6 hours. After 6 hours, supernatant media were collected and TNFα measured by ELISA. (B) BIRB 796 and dasatinib inhibit LPS-induced TNFα production in Fancc-deficient murine macrophages. Primary murine BM-derived macrophages obtained from Fancc^−/− and wild-type mice were maintained in control medium (plus DMSO) or treated for 6 hours with BIRB 796 (500nM) or dasatinib (500nM), followed by stimulation with LPS (1 μg/mL) for 24 hours. TNFα in conditioned media was measured by ELISA. (C-D) BIRB 796 and dasatinib inhibit R848- and LPS-induced TNFα production in FANCA-deficient human macrophages. Primary human CD14⁺ macrophages obtained from a normal age-matched volunteer and a FANCA-deficient patient were cultured for 6 hours in control medium (plus DMSO), BIRB 796 (500nM), or dasatinib (500nM), before stimulation for 24 hours with R848 (1μM; C) or LPS (0.1 ng/mL; D). TNFα in conditioned media was measured by ELISA.