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. 2015 Apr 1;9:113. doi: 10.3389/fncel.2015.00113

Figure 3.

Figure 3

FACS isolated Lgr5-positive cell divided and generated hair cells in culture. (A) Lgr5-GFP-positive and Lgr5-GFP-negative cells were isolated using FACS. (B,C) Immunostaining showed that among the FACS isolated Lgr5-EGFP-positive cells, 96.6% were GFP-positive, 96.7% were Sox2-positive, and none were Myo7a-positive. (D) Quantitative PCR showed that FACS isolated Lgr5-EGFP-positive cells had significantly higher Lgr5 expression, slightly higher Sox2 and Jagged1 expression, and significantly lower Brn3.1 expression compared to the Lgr5-EGFP-negative cells. (E,F) FACS isolated Lgr5-EGFP-positive cells generated significantly more colonies and Myo7a-positive cells than Lgr5-EGFP-negative cells. (G) Quantification of colony number generated from Lgr5-EGFP-positive and Lgr5-EGFP-negative cells. (H) Quantification of Myo7a-positive hair cells generated from Lgr5-EGFP-positive and Lgr5-EGFP-negative cells. (I) FACS isolated Lgr5-EGFP-positive cells mitotically generated hair cells in cell culture. (J) Quantification of EDU/Myo7a double positive hair cell number generated from Lgr5-EGFP-positive and Lgr5-EGFP-negative cells. *p < 0.05, **p < 0.01, ***p < 0.001, n = 3 in (D,G,H,J). Scale Bars: 20 µm.