Figure 3.

Gα₁₃ Arg-200 mutants induce NIH3T3 cell transformation in vitro.A, generation of NIH3T3 cell lines stably expressing the indicated Gα₁₃ proteins. Lentiviral particles for the expression of Gα₁₃ were generated in HEK293T cells and used to transduce NIH3T3 cells, followed by antibiotic selection. Lysates of each one of the cell lines were immunoblotted as indicated. Images were generated by splicing lanes from the same membrane, and the vertical dotted line indicates the position of the boundary between the two segments that were merged. B and C, Gα₁₃ Arg-200 mutants promote focus formation in NIH3T3 cells more efficiently than Gα₁₃ WT. Cells were seeded on plates and stained with crystal violet 10 days later. Images of a representative experiment are shown in B, whereas C shows the quantification of foci. Results are mean ± S. E. (error bars), n = 6. **, p < 0.01; ***, p < 0.001; n.s., not significant, analysis of variance with Tukey post hoc test.