Supplemental Fig. 1.

ATIP1 variant expression in HUVECs and anti-ATIP monoclonal antibody C1717 generation.

(A) The structural organization of the MTUS1 gene is quoted from the work of Di Benedetto et al. [20]. We designed primers in alternative exons used by the ATIP transcript variants as shown in Material and Methods. siATIP-N in exon 8 and siATIP-C in exon 17 indicates the siRNA region used to knockdown ATIP. (B) The Primer pairs, size (bp) and predicted ATIP transcript variants are listed. (C) The RT-PCR using the primers shown in (B) showed that the ATIP1 variant was expressed in HUVECs but not HeLa cells. (D) We selected the site of the immunogen to generate the anti-ATIP monoclonal antibody in the C-terminal region of mouse ATIP1 (327-426 a.a.), as indicated by both ends of the arrow. (E) C1717 detected Myc-tagged human ATIP1 transiently overexpressed in Hela cells as two bands of approximately 50 kDa. Overexpressed ATIP1 was also detected by anti-Myc antibody at the same size when using C1717. IB; immunoblot. Vehicle; cell lysate of pcDNA4 plasmid transfected to Hela cells. (F), (G) qPCR showed that endogenous ATIP1 was knocked down with siATIP-N or siATIP-C at the mRNA level. Values are the mean ± SEM for all three experiments. *p < 0.05.