Figure 2

Identification of putative surface markers of mDA NPCs. (A) Schematic of method. Using the 18a line, sample (top) was prepared by mDA induction medium for 14 days, and control (bottom) was induced in dual SMAD medium for 14 days. Both groups were incubated with LMX1 and FOXA2 antibodies and FACS purified. The resulting LMX1⁺FOXA2⁺ (sample) and LMX1^-FOXA2^- (control) populations were then processed through microarray to obtain the gene list. (B) The list of putative surface marker genes. The detailed list obtained through GO analysis can be found in the supplemental data. (C) Confirmation of RNA expression level of putative surface marker genes through qRT-PCR following MARIS. Following the 14 day-differentiation of BJ-RiPS, cells were fixed, stained with LMX1 and FOXA2 antibodies. These cells were then FACS purified into LMX1^-FOXA2^- and LMX1⁺FOXA2⁺ populations. The bar graph shows the relative expression of each putative surface marker gene in the LMX1⁺FOXA2⁺ population compared to the LMX1^-FOXA2^-population. (n = 2)