Figure 1
Figure 1. Gossypin inhibits both constitutive and inducible NF-κB activation. (A) Structure of gossypin. (B) Gossypin blocked NF-κB activation induced by TNF, H2O2, PMA, Lipopolysaccaride (LPS), interleukin-1β (IL-1β), okadaic acid (OA), cigarette smoke condensate (CSC). KBM-5 cells (2 × 106) were preincubated for 30 minutes at 37°C with 50 μM gossypin and then treated with TNF (0.1 nmol/L, 30 minutes), H2O2 (500 μM, 2 hours), PMA (25 ng/mL, 1 hour), LPS (100 ng/mL, 2 hours), IL-1β (100 ng/mL, 30 minutes), OA (500 nM, 4 hours), CSC (10 μg/mL, 1 hour). Nuclear extracts were prepared and assayed for NF-κB activation using EMSA. (C) Gossypin suppressed TNF-induced NF-κB in a dose-dependent manner. Human myeloid leukemia KBM-5 cells (2 × 106) were incubated with different concentrations of gossypin for 2 hours and then treated with 0.1 nmol/L TNF for 30 minutes. Nuclear extracts were prepared and assayed for NF-κB activation by EMSA. (D) Gossypin suppressed TNF-induced NF-κB in a time-dependent manner. Human myeloid leukemia KBM-5 (2 × 106) cells were incubated with 50 μM gossypin for the indicated time periods and then treated with 0.1 nmol/L TNF for 30 minutes. Nuclear extracts were then prepared and assayed for NF-κB activation by EMSA. (E) NF-κB consists of p50 and p65 subunits, and its binding to DNA is specific. Nuclear extracts from KBM-5 cells (2 × 106 cells/mL) left untreated or treated with 0.1 nm/L TNF were incubated at 37°C for 30 minutes with different antibodies or unlabeled NF-κB oligonucleotide probes and then tested for NF-κB activation, as described in “Materials and methods.” (F) Gossypin blocked TNF-induced NF-κB activation in human lung adenocarcinoma H1299 cells and embryonic kidney A293 cells that had been preincubated with 50 μM gossypin for 2 hours and 0.1 nmol/L TNF for 30 minutes. (G) Gossypin suppressed constitutive activation of NF-κB in BxPC3, U266, and 253JBV cells. Cells were incubated with 50 μM gossypin for 2 hours. Nuclear extracts were then prepared and assayed for NF-κB activation by EMSA. (H) Gossypin blocked various concentrations of TNF-induced NF-κB activation.

Gossypin inhibits both constitutive and inducible NF-κB activation. (A) Structure of gossypin. (B) Gossypin blocked NF-κB activation induced by TNF, H2O2, PMA, Lipopolysaccaride (LPS), interleukin-1β (IL-1β), okadaic acid (OA), cigarette smoke condensate (CSC). KBM-5 cells (2 × 106) were preincubated for 30 minutes at 37°C with 50 μM gossypin and then treated with TNF (0.1 nmol/L, 30 minutes), H2O2 (500 μM, 2 hours), PMA (25 ng/mL, 1 hour), LPS (100 ng/mL, 2 hours), IL-1β (100 ng/mL, 30 minutes), OA (500 nM, 4 hours), CSC (10 μg/mL, 1 hour). Nuclear extracts were prepared and assayed for NF-κB activation using EMSA. (C) Gossypin suppressed TNF-induced NF-κB in a dose-dependent manner. Human myeloid leukemia KBM-5 cells (2 × 106) were incubated with different concentrations of gossypin for 2 hours and then treated with 0.1 nmol/L TNF for 30 minutes. Nuclear extracts were prepared and assayed for NF-κB activation by EMSA. (D) Gossypin suppressed TNF-induced NF-κB in a time-dependent manner. Human myeloid leukemia KBM-5 (2 × 106) cells were incubated with 50 μM gossypin for the indicated time periods and then treated with 0.1 nmol/L TNF for 30 minutes. Nuclear extracts were then prepared and assayed for NF-κB activation by EMSA. (E) NF-κB consists of p50 and p65 subunits, and its binding to DNA is specific. Nuclear extracts from KBM-5 cells (2 × 106 cells/mL) left untreated or treated with 0.1 nm/L TNF were incubated at 37°C for 30 minutes with different antibodies or unlabeled NF-κB oligonucleotide probes and then tested for NF-κB activation, as described in “Materials and methods.” (F) Gossypin blocked TNF-induced NF-κB activation in human lung adenocarcinoma H1299 cells and embryonic kidney A293 cells that had been preincubated with 50 μM gossypin for 2 hours and 0.1 nmol/L TNF for 30 minutes. (G) Gossypin suppressed constitutive activation of NF-κB in BxPC3, U266, and 253JBV cells. Cells were incubated with 50 μM gossypin for 2 hours. Nuclear extracts were then prepared and assayed for NF-κB activation by EMSA. (H) Gossypin blocked various concentrations of TNF-induced NF-κB activation.

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