Figure 6.

The linc/circRNAs can improve iPSC reprogramming. A, effect of knockdown of the linc/circRNAs on formation of AP-positive iPSC colonies. MEFs were obtained from E13.5 embryos carrying a single copy of Dox-inducible expression of the four transcription factors. AP staining was performed at day 17 after doxycycline addition. Before Dox induction, 10,000 cells were placed in each well in a 6-well plate. Data are represented as mean ± S.D., n = 3. *, p < 0.001. B, DNA methylation analysis on the Dlk1-Dio3 locus in cell lines. C, expression of the Dlk1-Dio3 locus-encoded miRNAs in cell lines. The 2^−ΔΔCt method was used for analysis of the relative expression level of the miRNAs. The relative expression level of the miRNAs in control iPSCs was calculated as 1.00. Expression levels were normalized to control iPSCs. si-mix, cells transfected with a mix of siRNAs targeting on the linc/circRNAs; si-mix+Oct4 addback, cells transfected with mixture of siRNAs targeting on the linc/circRNAs and infected by retroviral Oct4. Each shape represents a cell line. D, images of chimeras obtained from si-mix+Oct4 addback iPSCs and its F1 progeny resulting from germline transmission. *, pup derived from the germline-competent iPSCs.