Fig. 5
From: Targeting the Akt–EphA2 axis and cell–cell adhesion enhances anoikis sensitivity in cancer cells
Tpm2.1 expression downregulates the Akt and EphA2 pathways in MDA cells under suspension growth conditions. (A) Western blot showing the expression of phospho-Akt (T308) and total Akt in MDA-EV and MDA-Tpm2.1 cells cultured under suspension and attached conditions for 5 days. (B,C) Quantification of phospho-Akt (T308), N=3 (B), and total Akt levels, N=2 (C), in MDA-EV and MDA-Tpm2.1 cells under suspension and attached conditions for 5 days, presented as Mean ± SD. Statistical significance was determined using two-way ANOVA. (D,E) Quantification of total apoptotic cells (%) presented as Mean ± SD for MDA-EV (D) and MDA-Tpm2.1 (E) cells after treatment with Akt inhibitor (5 µM) for 4 days under suspension and attached conditions. The Akt inhibitor was added at the time of seeding (0 h), 1 day, and 2 days after seeding, as indicated on the X-axis of panels D and E. Statistical significance was determined using two-way ANOVA followed by Šídák’s multiple comparisons test. (F) Western blot showing the expression of EphA2 in MDA-EV and MDA-Tpm2.1 cells cultured under suspension and attached conditions for 5 days. (G) Quantification of EphA2 levels in MDA-EV and MDA-Tpm2.1 cells under suspension and attached conditions for 5 days. N=1. (H) Quantification of total apoptotic cells (%) presented as Mean ± SD for MDA-EV and MDA-Tpm2.1 cells after treatment with EphA2 kinase inhibitor (1.5nM) for 4 days under suspension and attached conditions. Statistical significance was determined using an unpaired t-test followed by Welch’s correction and Holm-Šídák adjustment for multiple comparisons (α = 0.05). Uncropped versions of the Western blots are provided in Supplementary Figures 3 and 4. (*) for p value <0.05, (**) for p value < 0.01, (***) for p value < 0.001, (****) for p value <0.0001.
