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. 2023 Apr 12;299(5):104699. doi: 10.1016/j.jbc.2023.104699

Figure 7.

Figure 7

The p38-MK2-RSK-EphA2 pathway promotes U87-MG cell migration induced by TMZ.A, U87-MG cells were treated with DMSO or 100 μM temozolomide (TMZ) for 48 h. Whole-cell lysates were immunoblotted with primary antibodies against pS-EphA2, EphA2, pRSK, RSK1, RSK2, pp38, p38, pHSP27, HSP27 and β-Actin. B, U87-MG cells were transfected with siRNA against EphA2 (#1 or #2) or the negative control. After 5 h of transfection, cells were treated with DMSO or 100 μM TMZ for 72 h. Cell migration was observed using a time-lapse imaging system for 120 min. The accumulated distance of cell migration (μm) was calculated and shown in box and whisker plots. ∗p < 0.05 by the Tukey–Kramer HSD test. Whole-cell lysates were immunoblotted with primary antibodies against EphA2 and β-Actin. C–F, U87-MG cells were treated with DMSO or 100 μM TMZ for 72 h, then treated with DMSO, 10 μM BI-D1870 (C and E) or 10 μM MK2 inhibitor III (D and F) for 2 h. Whole-cell lysates were immunoblotted with primary antibodies against pS-EphA2, EphA2, pRSK, RSK1, RSK2, pHSP27, HSP27, and β-Actin (C and D). Cell migration was observed using a time-lapse imaging system for 120 min and the accumulated distance of cell migration (μm) was calculated and shown in box and whisker plots (E and F). ∗p < 0.05 by the Tukey–Kramer HSD test. DMSO, dimethyl sulfoxide; EphA2, ephrin type-A receptor 2; HSP27, heat shock protein 27; RSK, p90 ribosomal S6 kinase.