To verify loss of DALRD3, cell extracts were loaded onto BOLT 4–12% Bis-Tris gels (ThermoFisher) followed by immunoblotting onto PVDF membrane and probing with DALRD3 antibody (Proteintech, cat. No. 26294-1-AP, 1:1000 dilution) and anti-Actin C4 (EMD Millipore, cat. No MAB1501, 1:000 dilution). Expression of DALRD3 in stably-infected rescue cell lines were characterized by immunoblotting with the anti-FLAG M2 antibody (Sigma-Aldrich, F1804, 1:3333 dilution). Transient transfections of TWIN-Strep and FLAG-tagged proteins were also loaded onto BOLT 4–12% Bis-Tris gels, immunoblotted to PVDF membranes and probed with anti-TWIN-Strep (Genscript, cat. No. A01732, 1:1000 dilution) and anti-FLAG M2. NLuc immunoblots were probed with anti-FLAG, anti-GFP (Santa Cruz, cat. No. sc-9996; 1:100 dilution) and anti-Actin C4 antibodies. Image analysis of immunoblots were performed using Image Studio software (Li-Cor).
