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. 2017 May 16;151(4):474–480. doi: 10.1111/imm.12745

Figure 4.

Figure 4

Activation of the Akt–CREB signalling axis by Stressin‐1. (a) An EMSA with a labelled p53‐responsive element probe and recombinant p53 was performed in the presence or absence of Stressin‐1 (lanes 1 and 2), and the presence or absence of the monoclonal anti‐p53 antibody PAb‐421 (lanes 3 and 4). Stressin‐1 did not directly modify the band shift or PAb‐421‐induced supershift of the labelled probe. (b) Lipopolysaccharide (LPS) ‐induced nuclear factor‐κB (NF‐κB) activity of RAW‐blue reporter cells was assessed in the presence or absence of Stressin‐1, indicating that Stressin‐1 inhibited NF‐κB activity, (c, d) Stressin‐1 induced rapid phosphorylation of Akt kinase, GSK‐3 and CREB within minutes both in RAW 264.7 macrophages (c) and in NIH3T3 cells (d). (e,f) Stressin‐1 strongly reduced the LPS‐induced activation of signal transducer and activator of transcription 1 (STAT1) and STAT3 in RAW 264.7 cells, as assessed after 6 hr (e) and 24 hr (f) of LPS incubation. Values given in (c–f) indicate the band intensity relative to baseline samples (untreated or at 0 min) after normalization to actin bands. **P<0·01.