Figure 1 from Novel Pretreatment Autoantibodies Correlate with Enfortumab Vedotin–Related Dermatologic Events in Patients with Advanced Urothelial Cancer

<p>IPs from radiolabeled cell extracts using the pilot cohort sera (<b>A</b>), and validation of ROCK2 (<b>B</b>) and TOM1L1 (<b>C</b>) antibodies in the index sera. <b>A,</b> IPs were performed using <sup>35</sup>S-methionine–labeled MCF7 cell extracts and patient sera from the pilot cohort (sera #2–7) or a healthy control serum (#1). The IPs were electrophoresed by 10% SDS-PAGE, and immunoprecipitated proteins were visualized by autoradradiography. Migration of molecular weight standards are marked on the left. <b>B,</b> Antibodies against ROCK2 in index serum #4 from the pilot cohort were confirmed by IP from MCF7 lysates, followed by blotting with an anti-ROCK2 mAb. As a positive reference, the input lysate (lane denoted “lysate, no IP”) was included. <b>C,</b> The presence of TOM1L1 antibodies in index serum #5 from the pilot cohort was confirmed by IP using <sup>35</sup>S-methionine–labeled TOM1L1 generated by <i>in vitro</i> transcription and translation (IVTT). As a positive control, an anti-FLAG IP was performed (the IVTT protein is FLAG-tagged). The data in (<b>B</b>) and (<b>C</b>) confirm the presence of ROCK2 and TOM1L1 antibodies in sera #4 and #5, respectively.</p>