Runx2-I Isoform Contributes to Fetal Bone Formation Even in the Absence of Specific N-Terminal Amino Acids
Figure 3
The neo transgene perturbed Runx2 expression.
(A) Expression of total Runx2, Runx2-I, and Runx2-II in Runx2-I-mutated mice. Expression was analyzed by quantitative real-time PCR using RNA prepared from E18.5 calvarial cells of wild-type (+/+), Runx2-Ineo/+ (neo/+) and Runx2-Ineo/neo (neo/neo) mice. The results were normalized against the expression of Gapdh. Data are representative of three independent experiments and are means ± SEM (n = 3/group); *p<0.05, **p<0.01. (B) PCR primers (arrows) were placed on Runx2 exons and the neo transgene. (C) Semiquantitative RT-PCR analysis of RNA prepared from E18.5 calvarial cells of wild-type (+/+) and Runx2-Ineo/neo (neo/neo) mice by using the primer set depicted in panel (B). Gapdh was simultaneously amplified as an internal control (lanes 9 and 10). (D) Schematic diagram of alternative splicing pattern for Runx2-neo fusion transcripts. (E) Sequences of the fusion transcripts and proteins. The nucleotide sequence shown in italics is from neo, and the asterisk indicates the stop codon in the reading frame. The amino acids corresponding to each codon, and their numbers in the sequence of each Runx2 isoform are also indicated.