Fig. 4

Analysis of the biological effects exerted by RET-G533C and RET-P1047S mutants. Plasmids encoding wild type or mutant RET variants (G533C and P1047S) were transfected into HEK293 cells and selected in puromycin. a. MTT assay was performed at different time points (24 h, 48 h, 72 h and 96 h) using empty pBabe plasmid as negative control and plasmid encoding RET C634R mutant as positive control. Values are shown as bar graphs and all results are the average of two independent experiments performed in triplicate, Error bars s.d.; n = 6; **p < 0.01; ***p < 0.001 compared with control. b. Colony formation assay was performed with HEK293 cells transfected with plasmids encoding wild type or mutant RET variants (G533C and P1047S). We used empty pBabe plasmid as negative control and plasmid encoding RET C634R mutant as positive control. Similar results were observed in three independent experiments