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[Preprint]. 2024 Dec 4:2024.12.03.626601. [Version 1] doi: 10.1101/2024.12.03.626601

Figure 4. 10-SLF induces the formation of a ternary complex between FKBP12 and FBXW7-R465C.

Figure 4.

a. Coimmunoprecipitation assays demonstrated that HA-FBXW7-R465C and HA-FBXW7-R465C/C466A coimmunoprecipitated with FLAG-FKBP12_NLS in the presence of 10-SLF and MG132. The bar graph represents quantification of the immunoprecipitated HA-FBXW7 compared to HA-FBXW7 in whole cell lysates (WCL). Data are presented as the mean values (n = 2 independent replicates). b. FKBP12 enrichment proteomic analysis revealed that HA-FBXW7-R465C and HA-FBXW7-R465C/C466A interacted with FLAG-FKBP12_NLS in cells treated with 10-SLF. c. A NanoBRET assay measuring the interaction between FKBP12 and FBXW7 indicated that 10-SLF induced this interaction with FBXW7-R465C, but not with FBXW7 WT. Data are presented as the mean values ± s.e.m. (n = 3 independent replicates). The statistical significance was evaluated through unpaired two-tailed Student’s t-tests, comparing cells treated with 10-SLF to DMSO. *P < 0.05, **P < 0.01, ***P < 0.001, and ns: not significant.