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. 2016 Jan 27;7:4. doi: 10.3389/fendo.2016.00004

Figure 6.

Figure 6

Effect of camel milk on ERK1/2 and Akt phosphorylation. HEK293 cells transiently co-expressing hIR–Rluc8 and Grb2–Venus were used for ERK1/2 and Akt phosphorylation using the HTRF®-based assay (A–D) and SDS-PAGE/western blot technique (E,F). First, time-course analysis of phospho-ERK1/2 (A) and phospho-Akt (B) upon stimulation with 100 nM of insulin at different times was performed and HTRF signals were measured as described in Section “Materials and Methods.” Next, cells were first pre-treated or not 30 min with camel milk and stimulated or not 5 min with 100 nM of insulin before HTRF signals were measured as indicated. For the western blot analysis, cells were first pre-treated or not 30 min with camel milk and stimulated or not 5 min with 100 or 1 μM of insulin before SDS-PAGE and western blot using a rabbit polyclonal antibody against phospho-ERK1/2 (Thr202/Tyr204) (E). The western blot signals were quantified and expressed in arbitrary units after normalization (F). The HTRF data are mean ± SEM of three (for Akt) and four (for ERK1/2) independent experiments performed in triplicate. The western blot data are representative of two experiments.