Figure 5.

Chimeric minigene assay and RNA immunoprecipitation to map the determinants of the alternative splicing of exon 3. (A–D) The results of the chimeric minigene assay. Chimeric minigenes of human and mouse TREM2 were inserted into the pEGFP-C1 vector. The black boxes and lines indicate mouse exons and introns, respectively. The red boxes and lines indicate human exons and introns, respectively. The arrows exhibit the primer set. RT-PCR products were resolved by polyacrylamide gels. Tukey’s test was adopted for statistical tests (n = 4). (E) Western blot analysis to confirm the immunoprecipitation of CELF2 using the anti-CELF2 antibody. In input fraction. (F) RT-PCR analysis of cDNA synthesized from CELF2-immunoprecipitated products. The primer set, indicated by the black arrows, was used to detect the binding of CELF2 to TREM2 RNA.