Journal: Frontiers in Immunology
Article Title: FMNL1 and mDia1 promote efficient T cell migration through complex environments via distinct mechanisms
doi: 10.3389/fimmu.2024.1467415
Figure Lengend Snippet: FMNL1 mediates T cell migration independently of Myosin-II. WT, FMNL1 KO, and mDia1 KO T cells were collected from donor mice, ex vivo activated, and differentially fluorescently labeled. (A) T cells were seeded onto 3μm transwell inserts with 50μM Blebbistatin or a vehicle control and 100 ng/mL CXCL10 in the bottom well. Percent migration is calculated as the number of cells counted in the bottom well normalized to a 20% loading control well. (B–F) T cells were embedded in 1.0 mg/mL collagen with increasing concentrations of para-nitro-Blebbistatin. (B) Representative maximum Z-projection images of WT (red) FMNL1 KO (green), and mDia1 KO (cyan) T cell migration through 1.0 mg/mL collagen matrices treated with either DMSO (top) or 10 μM para-nitro-Blebbistatin (bottom). Track lines show the path taken by the cells over 25 min. (C) Trajectory plots of the DMSO and 10 μM Blebbistatin-treated groups for all cells analyzed in (D–F) . (D–F) Quantification of mean track speed, average arrest coefficient, and mean squared displacement of cells tracked continuously for 10 min. Cells were treated with a DMSO control, 3 μM or 10 μM para-nitro-Blebbistatin. Data in (A) represents the mean +/- SEM of three independent experiments. Significance was determined by One-way ANOVA. Data in (D–F) represent the mean +/- SEM of at least 165 cells per condition pooled from three independent experiments. Significance was determined by Brown-Forsythe and Welch ANOVA tests with Games-Howell’s multiple comparisons test. ns, not significant, * = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001.
Article Snippet: Recombinant Mouse IP-10 (CXCL10) , Peprotech , 250-16.
Techniques: Migration, Ex Vivo, Labeling, Control
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