Embryonic and postnatal brains were prepared and immunostained (Flandin et al., 2010), or assayed by in situ hybridization (Jeong et al., 2008). Protocols can be found on our lab website http://physio.ucsf.edu/rubenstein/protocols/index.asp, with modifications for dual immuno/in situ fluorescence analysis described in the Supplementary Experimental Procedures.
