Article Title: Cryo-EM analyses reveal the common mechanism and diversification in the activation of RET by different ligands
Figure Lengend Snippet: ( A ) Interface between RET and GFRAL. An overall view of the complex structure is shown in the middle as a reference. ( B ) Interface between RET-CRD and GDF15. ( C ) Pull-down assays for the GDF15/GFRAL/RET ternary complex. Strep-tagged GFRAL bound to Streptavidin-conjugated beads was used to pull down GDF15 and RET. Streptavidin beads without Strep-GFRAL bound served as negative control. Mutations in either GFRAL or GDF15 abolish binding of RET. The results shown are representative of three biological repeats. ( D ) GDF15-induced ERK phosphorylation in HEK293 cells expressing full-length myc-tagged RET and GFRAL. Cells were treated with GDF15 at 10 nM for 15 min. ERK phosphorylation levels (pERK) were assessed by western blot. Expression levels of RET and GFRAL were monitored by anti-myc western blot. The results shown are representative of three biological repeats.
Article Snippet: Cell line ( Homo sapiens ) , HEK293 , ATCC , Cat# PTA-4488, RRID: CVCL_0045 , .
Techniques: Negative Control, Binding Assay, Expressing, Western Blot