The CARD11 p.Cys150Leu variant does not confer GOF properties to murine B cells. (A) Schematic representation of the retroviral vector used for transduction of murine B cells. Mutations are indicated. The experimental procedure is shown on the right. (B) The frequency of transduced cells was determined by flow cytometric analysis of GFP⁺ signals at day 0, day 1, and day 2. (C) The total number of GFP⁺ cells was determined at different time points during the culture period, and proliferation was assessed by normalization to the number of GFP⁺ cells at day 0. (D) Expression of CD25 (left) and CD86 (right) was determined in triplicates at day 1. Gate was set on GFP⁺ cells. Representative data from 2 independent experiments are shown. (E) NF-κB–induced luciferase activity was normalized to constitutive Renilla luciferase signal. The graph shows technical triplicates. All data shown are representative for data obtained in 2 independent experiments.