Figure 7. TCGA LYN mutations.

(A) MCF-7 cells stably transduced with GFP, LYN^WT, LYN^D189Y or other LYN mutants described in TCGA (E159K, K209N, A370T, and G418R) were plated in 10% FBS. Cell lysates were harvested and analyzed by immunoblot using the indicated antibodies. Densitometry analysis was performed using ImageJ; the ratio of P-LYN_Y507/V5 bands is shown below each cell line column. PAR, parental. (B) Cells described in A were incubated in serum-free media for 24 or 48 hours. Protein lysates were then prepared and analyzed by immunoblot using the indicated antibodies. Calnex, calnexin (an antibody used in the immunoblot analysis). (C) Cells described in A were cultured in Matrigel with or without 1 μM BKM120 or 1 μM fulvestrant. On day 15, colonies were stained with MTT and photographed with a Gelcount reader. The average size of each cellular structure was quantitated and expressed relative to untreated GFP controls (n = 3; *P < 0.05 vs. respective treatments of GFP controls, ^#P < 0.001 vs. respective treatment of LYN^WT cells). Scale bars: 500 μm.