Extended Data Fig. 5: Local recruitment of CAMs is required to restore capillary blood flow.
From: Niche-specific dermal macrophage loss promotes skin capillary ageing
a, Scheme of long-term serial imaging of capillary niche in Cx3cr1-GFP;R26-mTmG mice. b, Sequential revisits reveal progressive pruning of capillary niche over a 6-month period. Yellow arrowheads indicate capillaries that will undergo pruning. c, Quantification of capillary-associated macrophage coverage at 1 and 7 months of age (n = 3 mice; three 500 µm2 regions per mouse; Frequency of macrophage association with capillary segments was compared between capillaries fated for pruning and all capillaries at 1- and 7-month-old time-points by two-way RM ANOVA and Fisher multiple comparison tests; mean ± SD). d, Scheme of laser-induced capillary clot experiment. e, Sequential revisits of damaged capillary segment (red dashed lines) after laser-induced clot formation in Cx3cr1-CreERT2;R26-mTmG mice. Yellow lightning bolt indicates site of laser-induced capillary clot. Yellow arrowheads indicate extra-luminal vascular debris. f, Quantification of capillary-associated macrophage extension toward damaged niche at Day 1, 2 and 7 after laser-induced clotting (n = 50 CAMs, in 3 mice; CAM extension toward capillary damage was compared by two-way RM ANOVA and Tukey multiple comparison tests; mean ± SD). g, Sequential revisits of damaged capillary niche after laser-induced clot formation (yellow lightning bolt) in Cx3crgfp/+ and Cx3crgfp/gfp mice. CAMs (green), capillaries (red), RBC (white). Yellow arrowheads indicate extra-luminal vascular debris. h, Quantification of capillary-associated macrophage extension toward damaged niche at Day 7 after laser-induced clotting (n = 99 CAMs in Cx3crgfp/+ group; n = 65 CAMs in Cx3crgfp/gfp group; n = 3 mice in each group; CAM extension (Cx3crgfp/+ vs Cx3crgfp/gfp) was compared by two-way RM ANOVA and Tukey multiple comparison tests; mean ± SD). i, Quantification of capillary reperfusion at Day 1, 2 and 7 after laser-induced clotting (n = 20 clots in Cx3crgfp/+ group; n = 21 clots in Cx3crgfp/gfp group; n = 3 mice in each group; capillary reperfusion (Cx3crgfp/+ vs Cx3crgfp/gfp) was compared by two-way RM ANOVA and Tukey multiple comparison tests; mean ± SD). j, Quantification of capillary blood flow (n = 4 mice in each group; two 500 µm2 regions per mouse; capillary blood flow (Cx3crgfp/+ vs Cx3crgfp/gfp) was compared at 1 and 6 months of age by two-way ANOVA and Fisher multiple comparison tests; mean ± SD). Scale bar, 50 µm.
