Epsins Regulate Mouse Embryonic Stem Cell Exit from Pluripotency and Neural Commitment by Controlling Notch Activation

Figure 4

EPN1 and EPN2 silencing affects neuroepithelial cells polarization and neural rosette lumen size. (a) Nestin immunofluorescence analysis reveals impaired neuroectoderm polarization and neural rosette lumen size in 7 day-neuralized EPN1 and EPN2 KD 46C mESC cultures. (b, c) Immunofluorescence analysis and relative size quantification of N-cadherin^+ve lumens show that EPN1 KD and EPN2 KD induce a decrease in the percentage of large-lumen rosettes. (d) After 11 days of exposure to neuralizing conditions, EPN1 and EPN2 KD mESC cultures show an increase in βIII-tubulin^+ve and MAP 2^+ve neurons as scored by immunofluorescence analysis. Nuclei are counterstained with Hoechst 33258. All data are expressed as the ((a, d): biologically independent experiments; (b, c): biologically independent experiments). Statistical significance (unpaired -test): .

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