Fig 1: AirID-CRBN with thalidomide and IMiDs presents neo-substrate candidates in cells.a Schematic diagram of AlphaScreen-based biochemical assay for detecting IMiDs-dependent interaction between CRBN and neo-substrates. b In vitro interaction assay between CRBN and neo-substrate candidates. The interaction between biotin-labelled bls-CRBN and FLAG-GST-neo-substrate candidates in the presence of DMSO or 20 µM thalidomide (Th), lenalidomide (Le), pomalidomide (Po) or 5-hydroxythalidomide (5HT) was analysed using the AlphaScreen-based biochemical assay. All AlphaScreen signals mean raw luminescent signal in the AlphaScreen-based biochemical assay. Error bars denote the standard deviation (independent experiments; n = 3). c Immunoblot analysis of IMiD-dependent protein degradation of ZNF536, ZNF687 and ZMYM2. HEK293T cells expressing AGIA-ZNF536, Myc-ZNF687 or ZMYM2-AGIA and FLAG-CRBN were treated with DMSO, thalidomide (Th), lenalidomide (Le), pomalidomide (Po) or 5-hydroxythalidomide (5HT) for 16 h. The experiment was repeated three times independently with similar results. d Immunoblot analysis of endogenous ZMYM2 protein levels in IMR32 cells treated with DMSO, thalidomide (Th), lenalidomide (Le), pomalidomide (Po), 5-hydroxythalidomide (5HT) or CC-885 for 24 h. The experiment was repeated three times independently with similar results. e IMR32 and HEK293T cells were treated with DMSO or pomalidomide (Po) for 24 h and ZMYM2 mRNA expression levels were measured by quantitative RT-PCR. Relative mRNA expression was determined using the expression level with DMSO treatment. Error bars denote the standard deviation (biological replicates; n = 4). f Immunoblot analysis of ZMYM2 protein levels in IMR32 and HEK293T cells treated with DMSO or pomalidomide (Po) in the presence of DMSO, 5 µM MG132 or 1 µM MLN4924 for 24 h. g Immunoblot analysis of ZMYM2 protein levels in IMR32 cells treated with DMSO or pomalidomide (Po) for 48 h ZMYM2 band intensity was normalised to tubulin band intensity. All relative band intensities are expressed as fluorescent signals relative to that of DMSO. Error bars denote the standard deviation (biological replicates; n = 3) and P-values were calculated by one-way ANOVA with Tukey’s post-hoc test (*P = 0.031). Source data are provided as a Source data file.