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. 2015 Jul 21;4(9):980–992. doi: 10.5966/sctm.2014-0219

Figure 2.

Figure 2.

Confirmation of the ability of OSR1-GFP/SIX2-tdTomato double knock-in human inducible pluripotent stem cell lines to monitor SIX2 expression. (A): The flow cytometric analyses of tdTomato-positive cells on culture day 19 induced with or without the factors used in our previously reported differentiation protocol for the intermediate mesoderm [6]. The data from three independent experiments are presented as the mean ± SEM (n = 3). (B): The results of the reverse transcription-polymerase chain reaction analyses of the tdTomato-positive and tdTomato-negative cell populations isolated on day 19 shown in (A). (C): Immunostaining using antibodies against SIX2 on the tdTomato-positive and tdTomato-negative cells isolated on day 19. Alexa Fluor 647 was used as the secondary antibody to eliminate any overlap with the GFP or tdTomato signals. Scale bars = 50 μm. Abbreviations: APC, allophycocyanin; GFP, green fluorescent protein; Tx, treatment with factors described in [6].