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. Author manuscript; available in PMC: 2018 Sep 11.
Published in final edited form as: Dev Cell. 2017 Aug 31;42(5):498–513.e6. doi: 10.1016/j.devcel.2017.08.003

Figure 7. Monocyte motility and directionality in vivo do not require the Arp2/3 complex.

Figure 7

A) Allele combination used for in vivo experimentation. CX3CR1-Cre-ERT2 produced from the endogenous CX3CR1 promoter. Lox-Stop-Lox tdTomato expressed from the endogenous Rosa26 promoter. The Arpc2 allele as previously described. Tamoxifen induces LoxP recombination. B) Control (Arpc2+/−) or Arpc2−/− blood monocytes harvested after tamoxifen treatment. Scale = 10 µm. C) 0.3 mm diameter wounds were induced on day 0. tdTom+ monocytes were imaged the next day as they were recruited to the wound edge (arrow). D) Representative fields demonstrating tdTom+ Control and Arpc2−/− monocytes moving in situ toward a one day old wound over 15 minutes. Arrow denotes the direction of the wound. Individual cells are highlighted across the image series with colored arrows. Scale = 20 µm. E) Wind rose plots generated from endogenous Control (Arpc2+/−) or Arpc2−/− monocytes migrating toward one day old wounds in vivo. Data is from one representative experiment. 0°on the rose plot is the direction of the gradient. F) Forward Migration Index (FMI) of Control (Arpc2+/−) and Arpc2−/− monocytes in vivo plotted as mean ± 95% CI. Data table relates mean ± SEM of cell speed (V), FMI, d/T and total cells tracked (N). Quantitative measurements were pooled from 5 Control or 6 Arpc2−/− adult mice imaged one day after wounding. G) Velocity distribution of Control and Arpc2−/− monocytes from in vivo migration tracks (N = 234 or 382, respectively). Each point on the x-axis corresponds to a range of 50 microns per hour (0–50, 51–100, etc.). Y-axis: fraction of Control (cyan) or Arpc2−/− monocytes (red) migrating within given range. X-axis labels correspond to the upper bound of the 50 micron per hour range. H) FMI distribution of Control and Arpc2−/− monocytes (N = 234 or 382, respectively) from in vivo migration tracks. Each x-axis division corresponds to a range of 0.14, while the y-axis is the fraction of each population that falls within given range. The dotted vertical line divides monocytes with a positive FMI (to the right) that are capable of migrating directionally from those to the left (FMI ≤ 0) that do not. See also Figure S7, Movie S6.