96 well opaque white plate

To measure the GLuc activity in infected cell cultures, we used a microplate luminometer (Orion II; Berthold) with an injector system. Fifty microliters of culture supernatant was plated into a 96-well opaque white plate (Nunc) and, by using the injector, was mixed with 50 μl 1-μg/ml native coelenterazine (a 1-mg/ml stock in methanol [Sigma] diluted in PBS). Enzymatic activity was measured in the well mode with the following sequence: injection (50 μl), with no shaking, delay for 0 s, measurement at 10 s. The values were analyzed with GraphPad Prism (version 5.0) software after using the remove baseline function.

To quantify Gaussia luciferase (GLuc) activity in infected cell cultures, we used a microplate luminometer (Orion II; Berthold) with an injector system, as described before (22 (link)). Briefly, 50 µL cell culture supernatant was placed into a 96-well opaque white plate (Nunc) and was mixed with 50 µL of PBS containing 1 µg/mL native coelenterazine (Sigma) prior to quantification.