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. Author manuscript; available in PMC: 2010 Jun 5.
Published in final edited form as: Cell Stem Cell. 2009 May 7;4(6):513–524. doi: 10.1016/j.stem.2009.04.015

Figure 3. Generation of Klf4 and c-Myc transgenic NOD ES lines.

Figure 3

(A) Summary of ESC derivation efficiency from NOD and permissive strain control blastocysts. (B) Representative images for NOD blastocyst at embryonic day 3.5 and ICM outgrowths after plating the blastocysts on feeder cells. The cells were infected with the indicated viruses and passaged in mESM. (C) Representative images of NOD colonies after infection with c-Myc and Klf4 viruses at passage two (P2) and stable established NOD ES lines at P5. NOD ES 1M was derived after infections with Ubi-c-Myc and three lines were obtained following Ubi-Klf4 infection (NOD ES 2K is shown). (D) Transgene expression by RT-PCR in various NOD ES lines. MEFs infected with O, S, K, M viruses were used as positive controls. (E) Southern analysis of c-Myc and Klf4 viral integrations in representative NOD ES cell lines derived following viral infection. NOD MEFs are used as background controls. (F–G) Karyotype and cell cycle analysis of transgenic NOD ES lines. (H) Immuno-fluorescent staining of NOD ES lines for pluripotency markers. (I) NOD ES cell derived chimeras from NOD ES 1M and 2K cell lines were mated with NOD mice with albino offspring indicating germline transmission.