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. 2015 Nov 3;1:15031. doi: 10.1038/celldisc.2015.31

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Copyright © 2015 SIBS, CAS

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Generation of ICAHCI offspring by H19^Δ1-neo^ΔAG-haESCs. (a) Subclones of PGK-neo knockouts in H19^Δ1AG-haESCs by PCR. The primer pairs P5–P6 were used. (b) DNA sequence of PCR products amplified from the H19 gene of H19^Δ1-neo^ΔAG-haESCs. (c) Establishment of the H19^Δ1-neo^Δ1AG-haESC line after FACS enrichment for haploid cells. (d) Chromosome counting in H19^Δ1-neo^Δ1AG-haESCs showing normal haploidy. (e) The distribution of absolute chromosome numbers of H19^Δ1-neo^Δ1AG-haESCs under cell culture conditions. (f) Immunofluorescence staining of H19^Δand H19^Δ1-neo^Δ1AG-haESCs. Scale bar, 20 μM. (g) Expression of imprinted genes measured by quantitative reverse transcription PCR (RT-qPCR). AGH-OG-3 AG-haESCs were used as control. Error bars, ±s.d. n=3. **P<0.01. (h) PCR analysis of PGK-neo knockout in H19^Δ1-neo^Δ1SC mice.