Fig. 2

Verification and analysis of the Ats-1 interaction protein SDCBP. A. Ats-1 protein self-activating activity verification. 01, negative control. 02, positive control. 03, Ats-1-pGBKT7/pGADT7. (A-A) Growth on the SD/–Leu/–Trp (DDO) medium. (A-B) Growth on the SD/–Ade/–His/–Leu/–Trp (QDO) medium. The results in the figure were obtained from three independent replicate experiments. B. Transfer back verification test of potential interacting proteins with Ats-1 protein. (B-A) Growth on the DDO medium. (B-B) Growth on the QDO medium. (B-C) Growth on the SD/–Ade/–His/–Leu/–Trp/X-α-gal (QDO/X) medium. 01–31, additional details of potential interaction proteins are provided in Table S4. The results in the figure were obtained from three independent replicate experiments. C. Interaction between Ats-1 and SDCBP. The plasmids encoding Ats-1-His and SDCBP-Myc were co-transfected into the HEK293T cells. After 48 h, the cells were collected and the whole-cell lysates were immunoprecipitated with anti-His antibody (left) or anti-Myc antibody (right) and protein A/G agarose beads. Ats-1 and SDCBP were then detected with Western blotting assays using the antibodies against either the His or Myc tags. The blots were cut prior to hybridisation with antibodies during blotting. The relative intensities for the Western blots were semi-quantified using ImageJ software and are presented as bar graphs in Fig. S2, where the results in the figure were obtained from three independent replicate experiments