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Figure 1


Fig. 1. Inhibition of PI3Ks impairs EB formation. EBs were generated in the presence of 0, 5 and 10 µM LY294002 (LY) as described in Materials and Methods. (A) Examples of day 5 EBs, original magnification 100x (10x/0.4 objective lens was used). Bars, 200 µm. (B) On day 3 to day 6 samples of EBs were trypsinized and individual cells counted. Data shown are mean (± s.e.m.) from five independent experiments. (C) The total number of EBs formed were counted on day 6. Data shown are mean (± s.e.m.) from four independent experiments. (D) Protein extracts were prepared from ES cells (ES) and EBs on the days indicated and separated by SDS-PAGE. Immunoblots were probed with an antibody detecting phosphorylated serine residues Ser235 and Ser236 of the ribosomal S6 protein ({alpha}-P-S6). The same immunoblot was stripped and reprobed for SHP-2 to assess equal protein loading. *P<0.05, **P<=0.01, ***P<0.005.





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