Fig. 4
From: A solution for highly efficient electroporation of primary cytotoxic T lymphocytes
Our new recovery medium surpasses all other media in promoting the function of primary mouse CTLs. A Total internal reflection fluorescence microscopy (TIRFM) image of a mouse CTL transfected with CD81-SEP and cultivated with our new RM following the method described in Fig. 1. CTLs were seeded on anti-CD3ε coated coverslips to form an immunological synapse and to induce secretion, which is monitored for 10 minutes at 10 Hz. The yellow arrow shows a multivesicular body. Its fusion is displayed as a sequence of enlarged snapshots on the right. The time points of the displayed snapshots are indicated in seconds. Scale bar: 5 μm. B Fluorescent intensity variation during exocytosis of the multivesicular body indicated with the yellow arrow in (A). C Quantitative analysis of the proportion of secreting CTLs indicated as percentage of secreting cells relative to the total number of transfected cells. Data represents mean ± SEM. N = 4 independent experiments, n = 31, 57, 20, 36, 19 cells in the order of RM displayed on the graph, *p < 0.05, **p < 0.01, *** p < 0.001