Figure 2. Identification and structure of 2 FIP-2s and their expression. (A) Clones obtained by cDNA library screening and RACE studies. WIN-11 (0.35 kb), pX105-2 (1.8 kb), and clone #1 (2.2 kb) significantly overlapped; therefore, they have been submitted to DDJB and assigned accession no. AB050777. Clones #2 (0.9 kb) and #3 (3.0 kb) were completely overlapped; therefore, they have been submitted to DDJB and assigned accession no. AB069907. *WIN-11 displaying the greatest difference in mRNA expression between the wounded and healthy pulp tissues by reverse Northern analysis; , common region in rFIP-2A and rFIP-2B; , region of rFIP-2B different from that of rFIP-2A. , cDNA containing the specific region for rFIP-2A or rFIP-2B was cloned and used as a template for riboprobes of in situ hybridization. The procedure is described in detail in the Appendix. (B) Structure of rFIP-2s. Deduced amino acid sequences of rFIP-2s and hFIP-2 are shown. The signal peptide domain is boxed in green, the putative leucine-zipper domains are boxed in black, and the zinc-finger domain is boxed in blue. The putative bZIP motif is boxed in red. *Same amino acid as the sequence above; –, missing amino acid. (C) Expression of rFIP-2 mRNA. Complementary DNA was amplified by PCR with the primers as described below. The PCR products for rFIP-2A, B, and ß-actin underwent electrophoresis on a gel. The specific primers were designed: rFIP-2A sense, 5'-TGCCCAGCCAGCCTCCTACC-3'; rFIP-2B sense, 5'-ATCTCTGTGGCCGGACCTGTTACC-3'; and rFIP-2A and rFIP-2B antisense, 5'-CCACTTCGATTCCCACACTC-3'. For an internal control, specific primers for rat ß-actin were designed: sense, 5'-TTGTAACCAACTGGGACGATATGG-3'; and antisense, 5'-GATCTTGATCTTCATGGTGCTAGG-3'. Two independent PCRs were performed, and typical results are shown. Lane M, 100-bp DNA ladder. Relative signal intensity (each mRNA from/ß-actin mRNA) is shown in the lower panel. , healthy pulp; , wounded pulp.