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. Author manuscript; available in PMC: 2014 Feb 1.
Published in final edited form as: Nat Cell Biol. 2013 Jun 16;15(8):978–990. doi: 10.1038/ncb2784

Figure 7. IL-1 signalling regulates senescence.

Figure 7

(a) IMR90 were infected as indicated and proliferation measured by CV. (b) SA-β-Gal staining. Scale bar, 50 μm. (c) IF using the indicated antibodies. Data is a representative experiment. Source data for 2 independent experiments are provided in Supplementary Table S8. (d) IMR90 ER:RAS were infected as indicated and cell growth analysed by CV (right). qRT-PCR showing knockdown efficiency (left). Data are one representative experiment out of 2 independent experiments. (e) IMR90 ER:RAS cells were tretaed as indicated. BrdU incorporation and p21CIP1 expression were measured. Data are mean ± s.d., n = 3 independent experiments. p was calculated using Student’s t-Test. For BrdU, IL1R inh p = 5.30×10−5; Casp inh I p = 0.013; Casp inh II p = 0.013. For p21, IL1R inh p = 6.73×10−6; Casp inh I p = 1.42×10−5; Casp inh II p = 0.00047. Source data are provided in Supplementary Table S8. (f) Knockdown of IL1R prevents paracrine senescence. IMR90 cells infected and treated as indicated. After 10-14 days, cells were stained for SA-β-Gal. Data is representative experiment. Source data for 2 independent experiments are provided in Supplementary Table S8. (g-i) Inhibition of IL-1R reduces senescence and immune clearance in vivo. (g) NrasG12V or NrasG12V/D38A transposons and a transposase were co-delivered into mouse livers via hydrodynamic injection. Mice were treated with carrier or drugs for 12 days. (h) Quantification of Nras- (top), p21Cip1- (centre) and p16Ink4a- (bottom) positive cells on liver sections. Student’s t-test (n=5 mice per condition): carrier/ NrasG12V vs. IL1R/ NrasG12V, *P< 0.0150, Carrier/ NrasG12V vs. Inhibitor combination/ NrasG12V, **P< 0.002, p21: Carrier/ NrasG12V vs. IL1R/ NrasG12V, *P< 0.0341, Carrier/ NrasG12V vs. Inhibitor combination/ NrasG12V, *P< 0.0116, p16: Carrier/ NrasG12V vs. Inhibitor combination/ NrasG12V, *P< 0.01. 1, delivery of NrasG12V/D38A and treatment with vehicle. 2-7 delivery of NrasG12V. Treatments: 2, vehicle; 3, IL1R inhibitor; 4, CCR2 inhibitor; 5, VEGFR2 inhibitor; 6, TGFBR1 inhibitor; 7, combination of the 4 inhibitors. Data are mean ± s.e.m. (i) Representative pictures of H&E and IHC for Nras, p2Cip1 and p16Ink4a. Scale bar, 100 μm.