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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Cancer Res. 2018 May 29;78(15):4360–4369. doi: 10.1158/0008-5472.CAN-17-3124

Figure 4. CDK4/6 inhibition drives NF-κB-mediated upregulation of NGF, BDNF, and HGF.

Figure 4

(A–B) Abemaciclib (1.5 μM) treatment upregulates NGF, BDNF, and HGF expression. Shown is an immunoblot using antibodies specific for NGF, BDNF, and HGF. Protein expressions were quantitated and plotted (*P < 0.05; **P < 0.01; ***P < 0.0001; two-tailed t-test). mRNA levels of NGF, BDNF, and HGF in G88 were detected with qRT-PCR following abemaciclib treatment at different time points (**P < 0.01; ***P < 0.001; two-tailed t-test). (C) Abemaciclib (1.5 μM) increases NF-κB activity detected with a luciferase reporter assay at 24 hours of treatment (**P < 0.01; ***P < 0.001; two-tailed t-test). (D–E) TNF-α (10 ng/ml) treatment upregulates NGF, BDNF, and HGF. Shown is an immunoblot using antibodies specific for NGF, BDNF, and HGF. Protein expressions were quantitated and plotted (*P < 0.05; **P < 0.01; ***P < 0.0001; two-tailed t-test). mRNA levels of NGF, BDNF, and HGF in G88 were detected with qRT-PCR following TNF-α treatment at different time points (***P < 0.001; two-tailed t-test). (F–G) Silencing NF-κB with a specific siRNA reverses the upregulation of NGF, BDNF, and HGF following abemaciclib (1.5 μM) treatment. Shown is an immunoblot using antibodies specific for NGF, BDNF, HGF, and NF-κB p65. Protein expressions were quantitated and plotted (*P < 0.05; ***P < 0.0001; two-tailed t-test). mRNA levels of NGF, BDNF, and HGF were detected with qRT-PCR following 24 hours of abemaciclib (1.5 μM) treatment in the presence of control and RELA siRNA (*P < 0.05; ***P < 0.0001; two-tailed t-test). All values are mean ± SEM and from three biologically independent samples.