Fig. 4
Basal activation of RhoA by the different TGAT mutants. Boxplot showing the median basal YFP/CFP ratio of the DORA RhoA biosensor in HeLa cells. Cells transfected with the constitutive active (ca, n = 70) or non-binding (nb, n = 62) RhoA biosensor were co-transfected with an empty vector containing only RFP to keep expression levels equal between the different experimental conditions. Wild-type (wt) RhoA biosensor was transfected with an empty vector containing just RFP (control, n = 107), RFP-RhoGDI (n = 118), RFP-TGAT (+ 2-bromopalmitate) (n = 129), RFP-TGAT (n = 291), RFP-TGATΔ15 (n = 169), RFP-TGATC242S, C253S (n = 148), RFP-TGATC242S (n = 153) or RFP-TGATC253S (n = 150). Boxplot center lines represent the median values; box limits indicate the 25th and 75th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles; data points from individual cells are plotted as dots