Supplementary Figure 3: Activity of the Oct4-CreER; R26-lsl-EYFP lineage tracing system in brain-derived NSCs
From: Lineage conversion induced by pluripotency factors involves transient passage through an iPSC stage
(A) Flow cytometric analysis of Oct4-CreER; R26-lsl-EYFP NSC cultures derived from the brains of five independent E13.5 embryos. Tamoxifen was administered to pregnant mice at E8.5 and brain-derived NSCs were recovered at day E13.5. EYFP expression was examined at passage 3 of in vitro culture. OKSM-iNSC clone #2 served as a positive control for EYFP expression. The PE-Cy7 channel was used to control for autofluorescence. (B) Representative immunofluorescence images show that the Oct4-CreER; R26-lsl-EYFP brain-derived NSC lines #1-5 express the NSC markers Sox1 and Sox2. (C) Flow cytometry analysis for uninduced Oct4-CreER; R26-lsl-EYFP brain-derived NSCs to confirm faithful regulation of the Oct4-CreER allele. Brain-NSCs were isolated at E13.5 and 4-OHT was added after 2 passages before performing flow cytometry analysis for EYFP. Note that no labeled cells were detected with or without 4-OHT in brain-NSCs of this genotype, indicating specificity of the Oct4-CreER allele. The PE-Cy7 channel was used to control for autofluorescence.
