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. Author manuscript; available in PMC: 2017 Feb 15.
Published in final edited form as: Nat Cell Biol. 2016 Aug 15;18(9):979–992. doi: 10.1038/ncb3397

Figure 2. Dynamic canonical NOTCH1 signalling is responsible for reciprocal regulation of TGF-β ligands and pro-inflammatory cytokines during senescence.

Figure 2

(a) Time series analysis of cell surface NOTCH1 expression during RIS in IMR90 cells by flow cytometry. Values are means relative to d0 ± SEM from 3 independent experiments. (b and c) Time course of protein expression by immunoblotting during RIS (b) or DDIS (c). (d) ER:HRASG12V IMR90 cells, expressing dnMAML1-mVenus or matched control, were incubated with or without 4OHT for 3 days and analysed for expression of indicated mRNA and proteins by qRT-PCR and immunoblotting respectively; n = 5 biologically independent experiments for TGFB1 and IL1B, n = 4 biologically independent experiments for IL1A; unpaired T-test. (e) ER:HRASG12V IMR90 cells, expressing a doxycycline-inducible N1ICD-FLAG construct (TRE-N1ICD) were analysed after 6 days treatment with 4OHT with or without doxycycline at indicated concentrations from d3 by qRT-PCR and immunoblotting; n = 6 biologically independent experiments for all conditions (except 4OHT / 1 μM Doxy where n = 5); unpaired T-test. Values are mean ± SEM; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. Statistics source data for a, d & e are provided in Supplementary Table 2.