Gating and quality control of flow cytometry. (A) GFP expression on two TMD8 BCR transductants in TKO with high and low transduction efficiency showing gating for BCR-expressing cells. Histograms show MFI of GFP. (B) GFP expression on two Karpas 422 BCR transductants in TKO with high and low transduction efficiency showing gating for BCR-expressing cells. Histograms show MFI of GFP. (C) Distribution of GFP-positive cells, GFP MFI, and calculated BCR signaling strength of all samples among all experiments. Each dot represents one measurement. Horizontal lines indicate the median. (D) Correlations of GFP MFI with transduction efficiency and with BCR signaling strength. Transduction efficiency is correlated with GFP MFI. BCR signaling strength is not correlated to GFP MFI. Transduction efficiency and BCR expression do not influence the calculation of autonomous signaling strength of the BCR.