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. 2013 Sep 30;110(44):17802–17807. doi: 10.1073/pnas.1312062110

Fig. 5.

Fig. 5.

Exogenous anti-FGF4 antibodies cause retained expression of pluripotency markers when added to the three independent ES cell lines. (A) Oct4–ΔPE-GFP ES cells grown in differentiation medium for 7 d in the presence or absence of purified αFGF4_A antibody (20 μg/mL) and compared with PDO3 control (magnification 20×). (B) Flow cytometry of Rex1-GFPd2 ES cells after 3 d of differentiation in the presence of αFGF4_A and compared with PDO3 control. (C) Flow cytometry of Nanog-GFP ES cells after 3 d of differentiation in the presence of αFGF4_A and compared with PDO3 control. (D) Percentages of cells expressing Rex1-GFP after 3 d of differentiation in the presence or absence of the indicated purified anti-FGF4 antibodies (20 μg/mL), negative (α-desmin and Fc only) and positive (PDO3) controls. The bars represent the average of triplicates of one flow cytometry experiment ± SD. (E) The histogram shows the fraction of Rex1-GFP+ cells (normalized to PDO3 response and using “minus antibody” as 0) from three independent experiments as described in D. The bars represent the average of triplicates of three separate flow cytometry experiments ± SD The asterisk denotes statistical significance. P = 0.0017 for αFGF4_A vs. antidesmin. In addition, P = 0.004 for αFGF4_C vs. α-desmin.