Extended Data Fig. 3: Interaction of the N-terminal part of ANGPTL4 with Syndecan-4 protects dermal microvascular endothelial cells from meningococcal infection.

a-d, HDMEC either uninfected or infected with N. meningitidis 2C4.3 for 30 min were treated with 1 μg mL⁻¹ of rhANGPTL4 or vehicle for 2h30. a, Cells were fixed and stained for cellular actin, VE-cadherin and DAPI and analysed by fluorescent microscopy (scale bars, 50 μm). b, c, Quantifications of VE-cadherin enrichment at intercellular junctions (b) and of intercellular spaces area (c) were performed with ImageJ software (n = 15 fields from n = 2 independent experiments). Error bars show mean ± s.e.m.; Ordinary one-way ANOVA Tukey’s multiple comparison test ****P < 0.0001; NS, not significant (NS1 = 0.994, NS2 = 0.9999). d, Infected HDMEC treated with 1 μg.mL⁻¹ of rhANGPTL4 or vehicle for 2 h and 30 min were lysed and the number of adherent bacteria was quantified (CFU.ml⁻¹). Error bars show mean ± s.d. from n = 2 independent experiments performed in duplicate; Unpaired t-test (two-tailed) NS, not significant (NS = 0.9357). e,f, Uninfected or infected HDMEC were treated with 1 μg.mL⁻¹ of rhANGPTL4, rhANGPTL3, rhANGPTL8 or vehicle for 2 h and 30 min. Quantifications of VE-cadherin enrichment at intercellular junctions (d) and of intercellular spaces area (e) were performed with ImageJ software;. Error bars show mean ± s.e.m (n = 14 fields from n = 2 independent experiments). Ordinary one-way ANOVA Tukey’s multiple comparison test ****P < 0.001; NS, not significant (NS1 = 0.9993, NS2 > 0.9999, NS3 = 0.5346, NS4 = 0.9221). g, Extended data of the images shown in Fig. 2a (scale bars, 50 μm). h, mRNA levels of SDC4 in uninfected and infected HDMEC or HBMEC are represented as normalized values in transcripts per million (TPM). Error bars show mean ± s.e.m. from one experiment performed in triplicate.

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