Figure 1
Figure 1. BIRB 796 and dasatinib inhibit NF-κB/AP-1–dependent SEAP production in T-shFC cells. Mean values ± SD are shown. All P values were calculated using the Student t test. (A) BIRB 796 inhibits R848-induced SEAP gene expression in FANCC-deficient cells. THP1-Blue cells (THP1) or THP1-Blue cells stably expressing either control (nontarget) shRNA (T-shNT) or shRNA targeting FANCC (T-shFC) were cultured in medium alone (control) or medium with the TLR7/8 agonist R848 (30μM) alone, BIRB 796 (500nM) alone, or R848 (30μM) alone after a 6-hour pretreatment with BIRB 796 (500nM). Conditioned media were collected after 24 hours of R848 exposure and SEAP was quantified colorimetrically using QUANTI-Blue reagent. (B) Dasatinib inhibits R848-induced SEAP expression in FANCC-deficient cells. The experimental design was identical to that of 1A except that dasatinib (500nM) was used as the inhibitor. (C) BIRB 796 inhibits TNFα secretion in THP-1 cells. T-shNT and T-shFC were treated as in panel A with BIRB 796 (500nM) and/or R848 (30μM). Conditioned media were collected after 24 hours of incubation with R848 and TNFα was quantified by ELISA. Untreated cells or cells treated only with BIRB 796 did not produce detectable TNFα (indicated with an asterisk). (D) Dasatinib inhibits TNFα secretion in THP-1 cells. T-shNT and T-shFC cells were treated as in panel B with R848 (30μM) and dasatinib (500nM). Untreated cells or cells treated with dasatinib alone did not produce detectable TNFα (indicated with an asterisk). (E) FANCA-deficient THP-1 cells are hypersensitive to R848. THP1-Blue cells stably expressing shRNA targeting FANCA (T-shFA) and T-shNT cells were exposed to R848 (0-100μM) for 24 hours. Supernatant media were collected and SEAP was quantified colorimetrically using QUANTI-Blue reagent. At each dose of R848, T-shFA cells produced more SEAP than did T-shNT cells (P < .05). (F) BIRB 796 and dasatinib suppress the production of SEAP in both T-shFA and T-shNT cells. The design of these studies on T-shFA cells was identical to that of studies on T-shFC cells (A-B). (G) BIRB 796 and dasatinib suppress TNFα production in R848-stimulated T-shFA and T-shNT cells. The design of these studies on T-shFA cells was identical to that used with T-shFC cells (C-D).

BIRB 796 and dasatinib inhibit NF-κB/AP-1–dependent SEAP production in T-shFC cells. Mean values ± SD are shown. All P values were calculated using the Student t test. (A) BIRB 796 inhibits R848-induced SEAP gene expression in FANCC-deficient cells. THP1-Blue cells (THP1) or THP1-Blue cells stably expressing either control (nontarget) shRNA (T-shNT) or shRNA targeting FANCC (T-shFC) were cultured in medium alone (control) or medium with the TLR7/8 agonist R848 (30μM) alone, BIRB 796 (500nM) alone, or R848 (30μM) alone after a 6-hour pretreatment with BIRB 796 (500nM). Conditioned media were collected after 24 hours of R848 exposure and SEAP was quantified colorimetrically using QUANTI-Blue reagent. (B) Dasatinib inhibits R848-induced SEAP expression in FANCC-deficient cells. The experimental design was identical to that of 1A except that dasatinib (500nM) was used as the inhibitor. (C) BIRB 796 inhibits TNFα secretion in THP-1 cells. T-shNT and T-shFC were treated as in panel A with BIRB 796 (500nM) and/or R848 (30μM). Conditioned media were collected after 24 hours of incubation with R848 and TNFα was quantified by ELISA. Untreated cells or cells treated only with BIRB 796 did not produce detectable TNFα (indicated with an asterisk). (D) Dasatinib inhibits TNFα secretion in THP-1 cells. T-shNT and T-shFC cells were treated as in panel B with R848 (30μM) and dasatinib (500nM). Untreated cells or cells treated with dasatinib alone did not produce detectable TNFα (indicated with an asterisk). (E) FANCA-deficient THP-1 cells are hypersensitive to R848. THP1-Blue cells stably expressing shRNA targeting FANCA (T-shFA) and T-shNT cells were exposed to R848 (0-100μM) for 24 hours. Supernatant media were collected and SEAP was quantified colorimetrically using QUANTI-Blue reagent. At each dose of R848, T-shFA cells produced more SEAP than did T-shNT cells (P < .05). (F) BIRB 796 and dasatinib suppress the production of SEAP in both T-shFA and T-shNT cells. The design of these studies on T-shFA cells was identical to that of studies on T-shFC cells (A-B). (G) BIRB 796 and dasatinib suppress TNFα production in R848-stimulated T-shFA and T-shNT cells. The design of these studies on T-shFA cells was identical to that used with T-shFC cells (C-D).

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