Fig. 2: The dynamic process of NTEM in mouse ICD model.

A Left panel: image displaying neutrophil form cluster in HF region. Yellow dashed circles indicate aggregated neutrophils. Right panel: representative neutrophil and monocyte contact image in dermis near HF region. Green arrows indicate the contacting cells. B Software (Imaris, Bitplane) differentiated 3 types of neutrophils with pseudo colors. HF-TEM-neutrophil (yellow), inter-HF-TEM neutrophils (blue), non-TEM neutrophils (red). C RFP fluorescent intensity integrated density (intDen) in HF neutrophils and inter HF TEM neutrophils, n = 47. D Representative image displaying the locations of neutrophils, monocytes, and hair follicles. Red: neutrophil, green: monocyte, blue: HF keratinocytes, white dashed circles indicating the HF region, scale bar: 50 μm. The upper parts of the images showed fluorescence-tagged neutrophils, monocytes, and keratinocytes. The lower parts were images displaying neutrophils, monocytes, and HF regions tracked by software (Imaris, Bitplane). E Mean fluorescent intensity (MFI) of RFP over time in HF (n = 6) and inter-HF regions (n = 9). F Comparison of RFP MFI in HF region of 0 min versus 180 min, n = 6. Mean track speed (G) and arrest coefficient (H) of HF-TEM neutrophil (n = 48), inter-HF-TEM neutrophil (n = 29), and non-TEM neutrophil (n = 86). I Frequency of neutrophils and monocytes at different distances to the closest HF marginal, N_{(neutrophils)}= 61,822, N_(monocytes)= 21,555. Mean track speed (J) and arrest coefficient (K) of monocytes in HF (n = 29) and inter-HF regions (n = 20). The arrest coefficient ranges from 0 to 1, and a value close to 1 indicates that cells have stopped moving. All the data were collected from female mice for these experiments. Data are presented as mean ± SEM from two independent experiments. Statistical analysis was performed using a two-tailed unpaired Student’s t-test.