Fig. 1: Immunoprecipitation (IP) with the rabbit anti-ELMSAN1 antibody.

a ELMSAN1’s binding to the IGFBPL1 bait is strongly inhibited by the KDM2A CPN domain. ELMSAN1 is shown by a red dot. Values on a are calculated from data of the previous publication: (gel slices #1–4 of Fig. 5a; -CPN nuclear extract of 5c)¹⁶. Y axis shows residual binding level of proteins in the presence of the CPN domain by ratio (C_b+CPN/C_b-CPN), and X axis shows enrichment of proteins by fold [(C/SOC)_b-CPN/(C/SOC)_e-CPN] with the nuclear extract lacking the CPN domain. (C/SOC)_b-CPN and (C/SOC)_e-CPN are concentration of a protein bound to the bait and the protein in the nuclear extract, respectively. Data for Y < 1 are plotted. b, c IP of nuclear extract with a rabbit anti-ELMSAN1 antibody. The IP is visualized by IB. nr-IgG (normal rabbit IgG), r-αELMS rabbit (anti-ELMSAN1 antibody), nExt (nuclear extract input). An asterisk (*) indicates noise derived from IgG. This symbol (*) was used throughout this report to indicate noise. The host of the antibodies used for the IB is either rabbit (r) or mouse (m). d Knockdown of the ELMSAN1 gene. Three different siRNAs and one universal negative control were added to 10 nM in duplicate cultures, respectively. Efficacy of the knockdown is estimated by dividing the level of ELMSAN1 with that of α-tubulin. e Inhibition of the rabbit ELMSAN1 antibody by the 6x His-tagged epitope (epELM-5). Twenty micrograms of epELM-5 bound to PVDF, and the control (28C) bound to PVDF (see Supplementary Fig. 1) were incubated in the IB solution that has both the r-anti-ELMSAN1 antibody and a sample PVDF membrane. The red arrowhead indicates the band for which binding of the antibody was reduced. f Identification of ELMSAN1 with a mouse monoclonal anti-ELMSAN1 antibody. PAGE separated immunoprecipitates were transferred to PVDF and probed with the rabbit (left) and the mouse monoclonal anti-ELMSAN1 antibody (right). Arrow (<) shows unidentified bands detected with the mouse antibody in the nuclear extract.