Figure 10.

GCaMP6S functions as an efficient and specific sensor for Ca²⁺ flux in Cas9 Hoxb8 cell-derived DCs.Ccr7^+/+ and Ccr7^−/− Cas9-Hoxb8 cells were transduced with a retrovirus expressing dTom and the Ca²⁺ sensor GCaMP6S. Successfully transduced cells were sorted based on the expression of dTom and subsequently differentiated to mature DCs in the presence of GM-CSF followed by LPS treatment. GCaMP6S intensity was measured by flow cytometry. After recording a baseline for 45 s, cells were treated with 500 ng/ml CCL21 and signals were recorded for additional 240 s. Finally, 1 μg/ml of ionomycin was added and signals were acquired for another 60 s. Data are representative for three independent experiments.