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. 2021 Oct 30;22(21):11824. doi: 10.3390/ijms222111824

Figure 3.

Figure 3

AZD8055 up–regulates phosphorylation of ERK1/2 and p38 and independently downregulates phosphorylation of 4E–BP1. (a) HepG2 cells were treated with 2 μM AZD8055 for various time points and immunoblot analyses were performed to measure the activation of three MAPKs using specific antibodies for ERK1/2, p–ERK1/2, p38, p–p38, JNK, and p–JNK. (b–d) HepG2 cells were treated with 2 μM AZD8055 for various time points (b) or pre–treated with or without 100 nM refametinib (c) or 10 μM SB203580 (d) for 1 h, followed by 2 μM AZD8055 treatment as indicated. Immunoblot analyses were performed using specific antibodies for 4E-BP1, phosphorylated form of 4E–BP1 at Ser65 (p–4E–BP1), ERK1/2, p–ERK1/2, p38, p–p38, and GAPDH. Three isoforms of 4E–BP1 are indicated (α, β, and γ). α and β isoforms are hypophosphorylated, and γ isoform is hyperphosphorylated. Open arrowheads and closed arrowheads indicate ERK1 and ERK2, respectively.