Skip to main content
. 2011 Aug 4;6(8):e22607. doi: 10.1371/journal.pone.0022607

Figure 2. CRaf RBCRD detects endogenous RasGTP.

Figure 2

(A) HEK 293T cells were transfected with Venus-RBDCRD, eCFP-CAAX PM reporter (PMem), and mCherry-H2B to label nuclei. Scale bar is 20 µm and applies to all panels. (B) KRas protein levels were reduced in NCI-H727 cells using 2 different siRNA oligos against KRas or control oligo and then transfected with Venus-RBDCRD. Scale bar is 20 µm and applies to all panels. (C) Quantification of RBDCRD PM targeting in NCI-H727 with KRas siRNA. (D) Hec1A isogenic cell ines (Parental, RasWT/- and RasG12D/-) were transfected with Venus-RBDCRD and imaged live. RBDCRD PM targeting was then measured for each cell line. (E) Serum starved HEK 293T cells were stimulated with 100 ng/ml EGF, and imaged every 15 sec, PM targeting of RBDCRD was then measured.