Figure 7: Ets-1 expression induced by BRAF and MEK inhibitors is blocked by Usp9x inhibition. | Nature Communications

Figure 7: Ets-1 expression induced by BRAF and MEK inhibitors is blocked by Usp9x inhibition.

From: Usp9x regulates Ets-1 ubiquitination and stability to control NRAS expression and tumorigenicity in melanoma

Figure 7

(a) Expression levels of the indicated genes (Ets-1, Ets-2, GABPA and Dusp4) by RT-PCR in NRAS mutant (WM1366) cells treated with PD0325901 for 0–3 h. (b) Immunoblot of the indicated proteins in NRAS mutant (WM1366) cells treated with PD0325901 for the interval noted. (c) Expression levels of the indicated genes (Ets-1, Ets-2, GABPA and Dusp4) by RT-PCR in NRAS mutant (WM1366) cells treated with PD0325901 for the interval noted. (d) Immunoblot for the proteins indicated in NRAS mutant (WM1366) cells treated with PD0325901 as described. (e) Expression levels of the genes indicated (Ets-1, Ets-2, GABPA and Dusp4) by RT-PCR in BRAF mutant (SK-Mel29) cells treated with vemurafenib for the interval indicated. (f) Relative luciferase units (firefly/Renilla) from NRAS mutant (WM1366) cells expressing the NRAS promoter for 24 h and treated with PD0325901 (0.5 μM) as noted. (g) Immunoblot for the proteins indicated in control and Usp9x KD NRAS mutant (SK-Mel147) cells treated with PD0325901 as indicated. (h) Phase contrast images of control and Usp9x KD NRAS mutant (SK-Mel147) cells treated with PD0325901 for 48 h. (i) Annexin V assessment in control and Usp9x KD NRAS mutant (SK-MEL147) cells treated with PD0325901 (1 μM) for 48 h as indicated. (j) Immunoblot for the proteins indicated in control and Ets-1 KD NRAS mutant (WM1366) cells treated with PD0325901 as indicated.

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