Cell count was analyzed with Imaris automatic
spot tracking aided by manual verification, and
cell migration was analyzed and verified with
manual tracking in 3D views. The surface of
large blood vessels was created according to
dextran-Tritic signaling. Spots and tracks that
lasted less than 1 min were excluded from
analysis. Migratory cDC2s were defined as cells
with at least a 30-mm track length across the
imaging period. The mean imaging period for
both types of chimeras was 45 min (number of
movies analyzed: 25Arhgef1+/−Batf3−/−Cd11c-
YFP and 29Arhgef1−/−Batf3−/−Cd11c-YFP).
Image sequences were annotated in Adobe
PhotoShop when necessary and rendered as
movies with Adobe AfterEffect. All movies are
played back at 20 frames per second (fps)
unless indicated otherwise, and time stamps
are minutes:seconds.
Statistical analysis
Statistical analysis and graphing were per-
formed using Prism 9.0 (GraphPad). Two-tailed
Student’sttests were used to compare endpoint
means of different groups. In grouped analysis,
ordinary two-way analysis of variance (ANOVA)
was performed and thePvalues indicated are
from individualttests with Sidak’s multiple test
correction. When multiple comparisons were
being performed, ordinary one-way ANOVA with
Dunnett’s multiple comparisons was used.
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ACKNOWLEDGMENTS
We thank S. Coughlin forArhGEF1-andGna13-deficient mice and
R. Brink for HEL2x. We thank M. De Giovanni for advice regardingLiuet al.,Science 375 , eabi5965 (2022) 11 February 2022 12 of 13
RESEARCH | RESEARCH ARTICLE