Figure 4 | Cell Death & Disease

Figure 4

From: Cancer cell death induced by novel small molecules degrading the TACC3 protein via the ubiquitin–proteasome pathway

Figure 4

SNIPER(TACC3) physically interacts with APC/CCDH1 and increases the TACC3–APC/CCDH1 interaction. (a) Cell lysates were mixed with 100 μM of SNIPER(TACC3)-1, Me-BS or KHS108 and analyzed by a thermal shift assay. (b) Scheme of the experimental procedure to detect the SNIPER(TACC3)-mediated interaction of TACC3 and APC/CCDH1. (c) HT1080 cells were co-transfected with the expression vector of Flag-TACC3 and Myc-CDH1. After 40 h, cells were treated with 25 μM of MG132 in the presence or absence of 30 μM of SNIPER(TACC3)-1 for 3 h. Cells were lysed in immunoprecipitated (IP) lysis buffer and Flag-TACC3 was IP with an anti-FLAG antibody. The precipitates were eluted with 2 mM of KHS108, Me-BS or 4-OHT. Total cell lysates, immunoprecipitates after elution and eluted fractions were western blotted with the indicated antibodies. DMSO, dimethyl sulfoxide; Elution, eluted fraction; lysate, total lysate; ppt, immunoprecipitate after elution; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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