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. 2013 Jul 11;93(1):173–180. doi: 10.1016/j.ajhg.2013.05.021

Figure 2.

Figure 2

Stimulation of ELK Transcription in NIH 3T3 Cells Expressing RIT1 Germline Mutations

(A) The ELK1-GAL4 vector and the GAL4 luciferase trans-reporter vector were transiently transfected with various RIT1 germline mutations and activating mutations in BRAF and MAP2K1 in NIH 3T3 cells. c.1910T>A (p.Val637Glu) in mouse Braf corresponds to oncogenic c.1799T>A (p.Val600Glu) in human BRAF. Relative luciferase activity was calculated by normalization to the activity of a cotransfected control vector, phRLnull-luc, containing distinguishable R. reniformis luciferase.

(B) ELK1 transactivation in cells expressing p.Ser35Thr, identified in individuals with Noonan syndrome, and p.Ser35Asn, were examined. p.Ser35Asn corresponds to dominant-negative alteration p.Ser17Asn in RAS.

Results are expressed as the means of quadruplicate (A) and triplicate (B) samples. Error bars represent the SDs of mean values. Red bars indicate germline RIT1 mutations identified in Noonan syndrome. The following abbreviation is used: WT, wild-type. ∗p < 0.01 by t test.