Figure 4. PFKFB4 and ICMT both control AKT signaling activation in melanoma cells.

(A) Protein levels of pAKT T308, pAKT S473, AKT, and actin in MeWo cells transfected with siRNA targeting PFKFB4 or ICMT. (B) Normalized pAKT S473/actin levels in MeWo, MNT1, A375, or Lu1205 cells transfected with siRNA targeting PFKFB4 and analyzed as in A; one point represents one biological replicate. (C) Protein levels of pERK, ERK, and vinculin in MeWo cells transfected with siRNA targeting PFKFB4. (D) Protein levels of pAKT S473 in MeWo cells treated with different concentrations of 2DG for 24 h. (E) Average speed of MeWo cells transfected either with siControl+empty vector, siPFKFB4+empty vector, siPFKFB4+xenopus PFKFB4 wild-type, siPFKFB4+caAKT, and siPFKFB4+ca-PI3K. In (A, C, D, E): a representative experiment is shown, n > 3. Graphs show the mean ± SEM. P-values were calculated using the Mann–Whitney test. n.s. P > 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

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