The Efficiency of Direct Maturation: the Comparison of Two hiPSC Differentiation Approaches into Motor Neurons

Figure 4

Early neuronal differentiation markers. (a) Representative confocal fluorescence microscopy images showing Pax6 (green) staining in the nuclei of NPC and Tubb3 (red) staining in the axons of all three approaches. Cell nuclei were counterstained with DAPI (blue). (b) Downregulation of Pax6 was detected by qRT-PCR in approaches A and B ( independent experiments). See Figure 3 for all results. (c) Decreased Pax6 protein expression was observed in Western Blot, shown in representative bands ( independent experiments). Each bar represents the fold change Deviation (SD). (d) NPC and neurons of approach A and B show persistent Nestin (green) and Tubb3 (red) expression detected by immunofluorescence. Cell nuclei were counterstained with DAPI (blue). (e) Decreased Nestin expression was detected in approaches A and B by flow cytometry and (f) by Western Blot, shown in representative bands ( independent experiments). Each bar represents the fold change . All scale bars, 50 μm.