DEV02264 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. In situ hybridization. Tbx1 expression in wild-type (A) and Foxg1^–/– (B) embryos, and Foxg1 expression in wild-type (C) and Tbx1^–/– (D) embryos at E10.5. Tbx1 expression is evident in the otic vesicle (ov) and the pharyngeal apparatus of both wild-type and Foxg1^–/– embryos. Foxg1 expression is evident in the telencephalon and in the OV of wild-type and Tbx1^–/– embryos. Scale bar: 0.1 mm.

• Supplemental Figure 2 -

  Fig. S2. Quantitative real time PCR (qRT-PCR) analysis of wild-type, Tbx1-null (Tbx1^–/–) and Tbx1 hygro/hygro (Tbx1 h/h) E9.5 embryos. To minimize variability of gene expression in individual embryos, a pool of five 9.5 dpc embryos for each genotype was used to test the expression level of Tbx1. Primers from both the deleted region (exon 3) and non-deleted region (exon 9) were used to ensure there was no splicing across the deleted region in Tbx1^–/– mice. Total RNA was isolated using the RNeasy Protect Mini Kit (Qiagen), and used for the first-strand cDNA synthesis with SuperScript First-Strand Synthesis System for RT-PCR (Invitrogen). The PCR reactions were performed in the LightCycler apparatus (Roche) using LightCycler-FastStart DNA Master SYBR Green I (Roche). The housekeeping gene Gapdh was chosen for internal normalization. Specific primers used for PCR amplification were as follows: Tbx1, Ex9-F (5¢-AGTACACGTTGGCTGCTGC-3¢), Ex9-R (5¢-AGTTCGACCGTGACTCCGGA-3¢), BstBI-F (5¢ACAAATAACCAGGCACTGGC-3¢) and 2g1-R (5¢-TCTTCTTGGGGCTGTAGACT-3¢); Gapdh, Gapdh-F (5¢-TTCACCACCATGGAGAAGGC-3¢) and Gapdh-R (5¢-GGCATGGACTGTGGTCATGA-3¢).