Fig. 6

Up-regulation of the CEBPB signaling is critical for the pro-aging effects of IGF2 deficiency. A Western blot analysis of protein levels of CEBPB in IGF2^f/f and IGF2^f/fCre mice of saline control group and d-gal model group. The quantitative data are presented. n = 5–7 per group. B Western blot analysis of protein levels of CEBPB in AML12 cells transfected with sh-IGF2 or sh-NC and treated with d-gal, H₂O₂ or the PBS control. C–H AML12 cells were transfected with sh-NC + si-NC, sh-NC + si-CEBPB, sh-IGF2 + si-NC and sh-IGF2 + si-CEBPB, and treated with d-gal or H₂O₂. C The senescence of AML12 cells detected by SA-β-Gal staining. β-gal staining area analysis was shown. Scale bar, 50 μm. D Immunofluorescence analysis of γ-H2AX expression in AML12 cells. Fluorescence intensity analysis was shown. Scale bar, 50 μm. Western blot analysis to measure the senescence-associated genes P53, P21 and P16 in AML12 cells treated with d-gal (E) or H₂O₂ (F). RT‒qPCR analysis of mRNA levels of senescence-associated secretory phenotype genes IL-6, IL-1β, TNF-α and NF-κB1 in AML12 cells treated with d-gal (G) or H₂O₂ (H). Data are shown as means ± SEM. One-way ANOVA was used for comparison among multiple groups. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. All experiments were repeated three times independently