Fig. 3.
SYK is essential for pre-adipocyte proliferation and for adipocyte differentiation. Representative western blot of SYK protein expression (a), cell numbers of three independent experiments (b) and representative flow cytometry of three independent experiments of CFSE staining analyzed by flow cytometry (c) of brown CreERT2 Sykflox/flox and Sykflox/flox pre-adipocytes seeded at equal cell numbers (d0) following continuous 4-hydroxy tamoxifen (4-OHT) treatment beginning 3 days prior. d Representative flow cytometry of three independent experiments of primary brown pre-adipocytes 3 days after labeling with CFSE and addition of 5 μM Syk-i3. Oil red oil staining (ORO), phase contrast images, and mRNA expression 4 days post induction of differentiation of primary brown (e, f) and white (g, h) pre-adipocytes in the presence of 5 μM SYK-i3 in the induction media; data are representative of three independent replicates. Mean cell number + SEM (b) and fold induction vs. d0 + SEM (f, h). For b, two-tailed unpaired Student’s t test was performed (*P < 0.05, **P < 0.005 and ***P < 0.0005) and for f, h, two-tailed unpaired Student’s t test was performed on Log transformed data