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. 2021 Mar 8;137(25):3533–3547. doi: 10.1182/blood.2020007731

Figure 5.

Figure 5.

mTORC1 activates IL-19 transcription by promotion of NF-κB signaling. (A) Expression of p-NF-κB (Ser536) and p-IKK-β (Ser180/181) was detected by western blotting in TSC1-loss and control osteocytes. (B) The expression of NF-κB in the nucleus and cytoplasm of TSC1-loss and control osteocytes. β-Actin and histone H1 were used as cytoplasm and nuclear protein markers, respectively. (C) The expression of p-NF-κB (Ser536) and p-IKK-β (Ser180/181) were detected by western blotting in Rheb-loss and control osteocytes. (D) The expression of NF-κB in the nucleus and cytoplasm in Rheb -loss and control osteocytes. (E) The level of IL-19 mRNA in TSC1-loss osteocytes exposed to NF-κB inhibitor (pyrrolidine dithiocarbamate [PDTC]; 50 µM, 24 hours) measured by qPCR (mean ± SD; n = 3). (F) IL-19 mRNA levels were detected by qPCR in control osteocytes exposed to PDTC (50 µM, 24 hours) or (and) mTORC1 activator PA (50 µM, 24 hours) (mean ± SD; n = 3). Data are mean ± SD of 3 independent experiments. **P < .01; ***P < .001.