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. Author manuscript; available in PMC: 2017 Jan 1.
Published in final edited form as: Cell Signal. 2015 Oct 19;28(1):43–52. doi: 10.1016/j.cellsig.2015.10.004

Figure 6. Decreased activity of phosphomimetic mutant of LARG.

Figure 6

(A) LARG activity was assessed via serum response element (SRE) luciferase activity. HEK293 cells were transfected with expression vectors for GFP-LARG, GFP-LARG-N/CpNull, GFP-LARG-N/CpMim or control pEGFP, along with both SRE-luciferase and Renilla luciferase. SRE-luciferase activity was measured and normalized as described under Materials and Methods. The graph shows quantitation (average ± SEM, n=3) of multiple experiments. Statistical significance is represented as follows: LARG compared to N/CpNull (*, p<0.05, t-test); N/CpNull compared to N/CpMim: (**, p<0.01, t-test); and LARG compared to N/CpMim: (***, p<0.0001, t-test). (B) Pull-down of LARG using GST-G17A-RhoA. The indicated GFP-tagged LARG proteins were expressed in HEK293 cells. Cell lysates were prepared and active LARG was pulled down using GST-G17A-RhoA. LARG in the affinity pulldown (upper panel) and in the cell lysate (lower panel) was determined by immunoblotting with a GFP antibody. A representative blot is shown. The graph shows quantitation (average ± SEM, n=4) of multiple experiments, in which the amount of LARG in the GST-G17A-Rho pulldown was normalized by the total expressed LARG in the lysate. The asterisk indicates statistical significance (p<0.05, t-test) of the N/CpMim mutant compared to wt LARG and to the N/CpNull mutant.