Figure 4. Effect of Stim1/2 ablation on... | Rockefeller University Press

Figure 4.

$Effect of Stim1/2 ablation on the spreading of neutrophils. (A) Average tdTomato fluorescence area and circularity of WT and Stim1/2KO-flushed neutrophils plated on PLL during the 3-min recordings in Fig. 2 and relative changes in the fluorescence area of these cells (n = 127/134 cells in 7/7 recordings from three WT/Stim1/2KO mice pairs, Student’s paired one-tailed t test on mice pairs. Small and large symbols show data from individual cells and mouse average, lines show median cellular values). (B) DIC micrographs (left) and fraction of spread WT and Stim1/2KO-flushed neutrophils plated on PLL for ∼20 min at 37°C and at RT (right, n = 37/39 and 11/11 recordings from 3 WT/Stim1/2KO mice pairs, Mann–Whitney U test. Student’s paired two-tailed t test on mice pairs). See Video 6 and Video 7. (C) TIRF micrographs of flushed neutrophils labeled with the PM dye CellMask adhering to PLL-coated glass (left, Video 8 and Video 9) and change in the size of neutrophil footprints after PLL contact (right, n = 25/25 graphed and 75/41 analyzed cells from four WT/Stim1/2KO mice pairs, two-tailed Welch’s t test on cell values).$

Effect of Stim1/2 ablation on the spreading of neutrophils. (A) Average tdTomato fluorescence area and circularity of WT and Stim1/2^KO-flushed neutrophils plated on PLL during the 3-min recordings in Fig. 2 and relative changes in the fluorescence area of these cells (n = 127/134 cells in 7/7 recordings from three WT/Stim1/2^KO mice pairs, Student’s paired one-tailed t test on mice pairs. Small and large symbols show data from individual cells and mouse average, lines show median cellular values). (B) DIC micrographs (left) and fraction of spread WT and Stim1/2^KO-flushed neutrophils plated on PLL for ∼20 min at 37°C and at RT (right, n = 37/39 and 11/11 recordings from 3 WT/Stim1/2^KO mice pairs, Mann–Whitney U test. Student’s paired two-tailed t test on mice pairs). See Video 6 and Video 7. (C) TIRF micrographs of flushed neutrophils labeled with the PM dye CellMask adhering to PLL-coated glass (left, Video 8 and Video 9) and change in the size of neutrophil footprints after PLL contact (right, n = 25/25 graphed and 75/41 analyzed cells from four WT/Stim1/2^KO mice pairs, two-tailed Welch’s t test on cell values).