HEK293, A375, and SW480 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS). Transient transfections were performed with TransFectin reagent (Bio-Rad, #1703352) or jetPRIME (Polyplus). Primary antibodies used were mouse anti-Rluc antibodies (Chemi-Con, #MAB4400 versus Rluc-F[2] and #MAB4410 versus Rluc-F[1]), mouse anti–HA-tag (Covance, #MMS-10P), mouse anti-FLAG (Sigma-Aldrich, #F3165), mouse anti-V5 (Invitrogen, #R9302), mouse anti-BRAF [Santa Cruz Biotechnology, sc-5284 (#F-7) and sc-166 (#C-19)], rabbit anti-CRAF (Cell Signaling Technology, #9422), mouse anti-RAS (Thermo Fisher Scientific, #MA1-012X), mouse anti-GFP (green fluorescent protein) (Roche, #11814460001), mouse anti-MEK1/2 (Cell Signaling Technology, #4684S), rabbit anti–P-MEK1/2 (Ser217/Ser221) (Cell Signaling Technology, #9154), rabbit anti–P-ERK1/2 (Cell Signaling Technology, #9101), rabbit anti-ERK1/2 (Cell Signaling Technology, #4696S), and mouse anti-PKA RIIβ (BD Biosciences, #61062).

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