Figure 2.
Role of NF-κB1 and MEK1 in KIT D816V-dependent expression of TNF. (A) Signaling pathways downstream of KIT D816V and applied pharmacological inhibitors. (B) Effects of different pharmacological inhibitors on TNF expression. HMC-1.2 cells were treated with PD98059 (50 μM), BEZ235 (1 μM), or TPCA-1 (20 μM) for 24 hours and TNF mRNA expression levels were measured by qRT-PCR. (C-D) HMC-1.2 cells were treated with ulixertinib (30-1000 nM) (C) or binimetinib (30-1000 nM) (D) for 5 hours and TNF expression was determined as described above. (E-F) HMC-1.2 cells were transduced with doxycycline-inducible RNAi against NF-kB1 (E), MAP2K1/MEK1 (F), or a nontargeting control (NTC). After induction with doxycycline (1 μg/mL) for 48 hours, TNF mRNA expression levels were measured by qRT-PCR. Results represent the mean and standard error of the mean of 3 independent experiments. ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001.

Role of NF-κB1 and MEK1 in KIT D816V-dependent expression of TNF. (A) Signaling pathways downstream of KIT D816V and applied pharmacological inhibitors. (B) Effects of different pharmacological inhibitors on TNF expression. HMC-1.2 cells were treated with PD98059 (50 μM), BEZ235 (1 μM), or TPCA-1 (20 μM) for 24 hours and TNF mRNA expression levels were measured by qRT-PCR. (C-D) HMC-1.2 cells were treated with ulixertinib (30-1000 nM) (C) or binimetinib (30-1000 nM) (D) for 5 hours and TNF expression was determined as described above. (E-F) HMC-1.2 cells were transduced with doxycycline-inducible RNAi against NF-kB1 (E), MAP2K1/MEK1 (F), or a nontargeting control (NTC). After induction with doxycycline (1 μg/mL) for 48 hours, TNF mRNA expression levels were measured by qRT-PCR. Results represent the mean and standard error of the mean of 3 independent experiments. ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001.

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