FIGURE

Figure 3—figure supplement 3.

ID

ZDB-FIG-220430-60

Publication

Peng et al., 2022 - Proper migration of lymphatic endothelial cells requires survival and guidance cues from arterial mural cells

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Figure 3—figure supplement 3.

Gating strategies of fluorescence activated cell sorting (FACS) and gene analysis of chemokine receptor on sorted lymphatic endothelial cell (LEC).

(A) Illustration of FACS and qPCR analysis on 3 days post fertilization (dpf) Tg(prox1a:TagRFP; fli1a:nGFP) embryos. (B) (Left) Gating strategy for FACS sort of Tg(prox1a:TagRFP; fli1a:nGFP) as described in Figure 3—figure supplement 3A. Sorting was performed on all singlet, alive cells according to their expression of DsRed (red, 561 nm) and GFP/ YFP (green, 488 nm). (Right) RT-qPCR of cxcr4a, cxcr4b, and ackr3b expression in trunk LECs at 3 dpf. Graph represents gene expression relative to geometric average of kdrl and β-actin from three biological repeats (mean ± SEM). Unpaired two-tailed Student’s t-test or Mann-Whitney test was used. *p < 0.05, **p < 0.001. Ns, no significance. (C) Gating strategy for FACS sort of Tg(flt1:YFP) and Tg (abcc9:Gal;UAS:GFP) as described in Figure 3D. Sorting was performed on all singlet, alive cells according to their expression of GFP/ YFP (green, 488 nm).

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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