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. 2019 Dec 16;161(2):bqz030. doi: 10.1210/endocr/bqz030

Figure 6.

Figure 6.

LepRb directly interacts with Fyn. (A) Schematic of LepRb–BioID experiment with representative Western blot of V5-Fyn immunoprecipitated from HEK293T cells expressing the designated BioID constructs and V5-Fyn. (B) Schematic of BioID-Fyn experiment with representative Western blot of LepRb-V5 immunoprecipitated from HEK293T cells expressing LepRb-V5 and either BioID-Fyn or a soluble BioID (BirA*). (C) Quantification of IP biotinylated LepRb-V5 intensity normalized to total LepRb-V5 intensity in the same lane (n = 3). (D) Representative Western blot of LepRb-myc immunoprecipitated from HEK293T cells stimulated with leptin (50 nM, 2 hours) and expressing LepRb-myc and V5-Fyn. (E) Quantification of coimmunoprecipitated V5-Fyn intensity normalized to immunoprecipitated LepRb-myc intensity from the same lane (n = 3). All BioID experiments were stimulated with biotin (50 µM) at the time of transfection. All experiments were repeated in 3 independent culture preparations and expressed as the mean ± SEM, **P < 0.01 compared to control, Abbreviations: IP, immunoprecipitate; Strep, streptavidin; WB, western blot.