Fig. 2: EndoBind allows for the detection of the interaction of endogenous KRAS and CRAF in PATU8988T cells.
From: EndoBind detects endogenous protein-protein interactions in real time
a EndoBind signal in PATU8988T-SmBKRAS-LgBCRAF cells or parental PATU8988T cells treated with GDC0879 for 1, 7, or 24 h (mean ± s.d., n = 3 wells each, representative experiment). Data were normalized to the control treatment (DMSO). b EndoBind signal after knockdown of KRAS or CRAF in PATU8988T-SmBKRAS-LgBCRAF cells and treatment with GDC0879 (mean ± s.d., n = 3 for siKRAS, mean of n = 2 for siCRAF). Signals were normalized to cells transfected with a non-targeting siRNA. Protein levels after transfection of the indicated amount of siRNA were quantified using the Jess system (mean, n = 2). c EndoBind signal in PATU8988T-SmBKRAS-LgBCRAF cells after treatment with the RAF-inhibitors GDC0879, LY3009120, NVP-LHS533, and the MEK-inhibitor Selumetinib. The same plate was measured at the indicated time points (mean ± s.d., n = 3 wells each, representative experiment). Data were normalized to the control (DMSO) treated sample. d EndoBind signal in PATU8988T-SmBKRAS-LgBCRAF cells after treatment with the Hsp90 inhibitor NVP-AUY922 and GDC0879 (mean ± s.d., n = 3 wells each, representative experiment). Data were normalized to the control (DMSO) treated sample. e EndoBind signal in PATU8988T-SmBKRAS-LgBCRAF cells after treatment with the Hsp70 inducer AEG3482 and GDC0879 (mean ± s.d., n = 3 wells each, representative experiment). Data were normalized to the control (DMSO) treated sample. f Distribution of EndoBind signal across the 35 plates (1536-well plates) of a 50 k compound screen in PATU8988T-SmBKRAS-LgBCRAF cells after treatment with NVP-AUY922 (active control), DMSO (neutral control), or sample compound in the presence of GDC0879. Data were normalized to the neutral control (DMSO) on each plate.
