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. 2016 Oct 25;6:35947. doi: 10.1038/srep35947

Figure 5. Restraint of Erk1/2 activation through modulation of Sos1-Grb2 association by induced Dab2 during adipocyte differentiation.

Figure 5

dab2 HET and CKO MEFs were subjected to adipose differentiation following the two-step protocol: 2 days in the “Induction Medium” followed by a time course in the “Differentiation Medium”. Grb2 and associated proteins were immunoprecipitated from Dab2 HET and CKO MEF cells on days 0, 1, 4, 9 of adipose differentiation. (a) Western blot analysis of Dab2, p-Erk1/2, Sos1, and Grb2 was performed on total cell lysates. (b) Western blot analysis of Dab2, Sos1, and Grb2 was performed on anti-Grb2 co-immunoprecipitation (co-IP) samples. (c) dab2 HET and CKO MEFs were first treated with “Induction Medium” for 2 days and then were cultured in the “Differentiation Medium” to mature into adipocytes in the presence or absence of MEK inhibitor U0216 for 4 days. Morphology of the cells was recorded, the cells were stained with Oil-Red O, and the adipocyte content was quantified. (d) The total cell lysates were analyzed by Western blot for markers including Dab2, phospho-Erk1/2, PPARγ, C/EBPα, and beta-actin. All Western blots compiled within a panel were derived from the same cell lysates and processed in an identical condition.