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. 2024 Jan 9;15:407. doi: 10.1038/s41467-024-44688-3

Fig. 6. Impact of LFA-1 activation blockade on degranulation, contact frequency and killing.

Fig. 6

a Flow cytometry analysis of LAMP1 and m24 expression on CD8+ T cells stimulated by P815 cells coated with the indicated concentrations of anti-CD3 Ab. Top raw displays CD8+ T cells treated with DMSO while bottom raw displays CD8+ T cells treated with 10 µM BIRT 377. One representative experiment among three independent experiments is shown. N = 20,000 cells for each condition. b In CD8+ T cells stimulated with P815 cells coated with 1 µg/ml anti-CD3 Ab, MFI of m24, LAMP1, TS2/4 and Hi-111 stainings depending on the treatment (DMSO or BIRT 377) applied to the cells. p values calculated by two-sided ratio paired t-test. Three independent experiments are shown. c Contact frequency between P815 cells coated with anti-CD3 Ab (1 µg/ml) and CD8+ T cells depending on the treatment (DMSO or BIRT 377) applied to the cells. The contact frequency is defined as the number of contacts divided by the number of live targets. Points represent the mean of three independent experiments and the error bars represent the SEM. The significance of the differences between the curves is determined at the 2-h and 6-h time points with two-sided unpaired t-tests. d Percentage of killing of the P815 cells coated with anti-CD3 Ab (1 µg/ml) depending on the treatment (DMSO or BIRT 377) applied to the cells. Killing values are calculated from the number of residual alive P815 cells at the considered time points. Points represent the mean of three independent experiments and error bars represent the SEM. The significance of the differences between the curves is determined at the 6, 12, 18 and 24-h time points with two-sided unpaired t-tests. Gating strategy exposed in Supplementary Fig. 5a. Source data are provided as a Source Data file.