Fig. 5: MCs promote graded expression of ICAM-1 and CXCL1 on pericytes. | Nature Communications

Fig. 5: MCs promote graded expression of ICAM-1 and CXCL1 on pericytes.

From: Neutrophil breaching of the blood vessel pericyte layer during diapedesis requires mast cell-derived IL-17A

Fig. 5

a–d TNF-stimulated or PBS-treated WT cremaster muscles were immunostained for MCs (avidin), pericytes (α-SMA) and ICAM-1 or CXCL1. a High magnification confocal image of a TNF-stimulated post-capillary venule (halved) showing enrichment of ICAM-1 (pseudocolour intensity) associated with pericytes (purple) in venule segment with or without MCs (cyan). White dashed line shows the limit of MC, scale bars, 5 µm. b, c Quantification of pericyte b ICAM-1 (n = 11 PBS no MC, n = 10 PBS < 3 MCs, n = 9 > 3 MCs, n = 9 TNF no MC, n = 12 TNF < 3 MCs, n = 8 TNF > 3 MCs, *p value = 0.0451, **p value = 0.0047, ***p value<0.0001, #p value = 0.0486) or c CXCL1 (n = 12 PBS no MC, n = 13 PBS < 3 MCs, n = 7 > 3 MCs, n = 15 TNF no MC, n = 15 TNF < 3 MCs, n = 7 TNF > 3 MCs, *p value = 0.0411, **p value = 0.0028, **p value < 0.0024, #p value = 0.0087) MFI in venules without, less than or more than 3 MCs. d–f Quantification of pericyte ICAM-1 MFI in 5 µm-wide consecutive regions (as exemplified in a) from a perivascular MC in d PBS and TNF treated WT mice (n = 18 PBS, 31 TNF perivascular MC regions, data pooled from five mice, *p value = 0.0366, ***p value = 0.003, ##p value = 0.0044), e PBS and TNF treated IL-17AKO mice (n = 36 perivascular MC regions, data pooled from five mice) or f in TNF treated mice in combination with ctrl mAb or anti-IL-17A mAb (n = 27 Ctrl mAb, 28 anti-IL-17A mAb perivascular MC regions, *p value = 0.0265, #p value = 0.0133); data pooled from four mice. g–i Quantification of pericyte CXCL1 MFI in 5 µm-wide consecutive regions from a perivascular MC in g PBS and TNF-treated WT mice (n = 31 PBS, 30 TNF perivascular MC regions, data pooled from five mice, *p value = 0.0322, ***p value = 0.003, ##p value = 0.0041), h PBS and TNF treated IL-17AKO mice (n = 41 perivascular MC regions, data pooled from five mice) or i in TNF treated mice in combination with ctrl mAb or anti-IL-17A mAb (n = 21 Ctrl mAb, 27 anti-IL-17A mAb perivascular MC regions *p value = 0.0248, #p value = 0.0448); data pooled from four mice. ICAM-1 and CXCL1 MFI were normalised to the most distal region from the MC (i.e., 15–20 µm). Mean ± SEM (each mouse represents one independent experiment). b–i two-way ANOVA followed by Sidak’s post-hoc test. *p < 0.05, **p < 0.01, ***p < 0.001 as compared to PBS or 20–15 region or as indicated by #p < 0.05, ##p < 0.01 (ns = not significant). Source data are provided as a Source Data file.

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