Tetraploid blastocysts were produced by electrofusion, and ES cells injected to make entirely ES-cell-derived embryos by tetraploid complementation (Nagy et al., 1993 (link)). Whole-mount X-gal staining of embryos was performed at E11.5 as described previously (Lettice et al., 2003 (link)), but on this occasion the staining was allowed to proceed at room temperature for between 1 and 18 h in a concentration of 300 µg/ml X-gal.
Wild-type mouse embryos were harvested at E11.5 and in situ hybridisation was performed with DIG-labelled gene-specific antisense probes as previously described (Hecksher-Sorensen et al., 1998 (link)).
Probes were generated for Lmbr1, Rnf32, Nom1 and Rbm33 by RT-PCR and cloned into the pBluescript vector (Agilent Technologies) (primers are listed insupplementary material Table S1 ). The Shh probe was kindly provided by Andy McMahon (Echelard et al., 1993 (link)) and the Cnpy1 and En2 probes by Jean M Herbert (Paek et al., 2012 (link)).
Wild-type mouse embryos were harvested at E11.5 and in situ hybridisation was performed with DIG-labelled gene-specific antisense probes as previously described (Hecksher-Sorensen et al., 1998 (link)).
Probes were generated for Lmbr1, Rnf32, Nom1 and Rbm33 by RT-PCR and cloned into the pBluescript vector (Agilent Technologies) (primers are listed in
Free full text: Click here
