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. 2016 Sep 26;36(20):2612–2625. doi: 10.1128/MCB.00191-16

FIG 6.

FIG 6

SPRED1 mutants found in Legius syndrome show decreased interactions with Gal-1 and B-Raf. (A and B) Interaction between mGFP-tagged SPRED1 mutants and mRFP–Gal-1 or mCherry–B-Raf was detected using FRET in HEK cells transiently expressing the constructs as indicated. Numbers inside the bars correspond to the total number of cells studied in each case. Error bars indicate the standard errors of the means. ***, P < 0.001. (C) SPRED1 membrane translocation FRET assay was conducted in HEK cells transiently coexpressing an mGFP-tagged SPRED1 mutant and mCherry-tagged wild-type SPRED1. The cells were treated with 0.5% DMSO (control cells), or starved for 5 h and stimulated with EGF (100 ng/ml, 10 min), or treated with sorafenib (50 μM, 2 h). Numbers inside the bars correspond to the total number of cells studied in each case. Error bars indicate the standard errors of the means. ***, P < 0.001; ns, nonsignificant.