Fig. 5: CRELD2 promotes angiogenic effects.

a, Exemplary immunoblots showing CRELD2 expression (normalized to TUBA) in HCAEC lysates (L) and conditioned supernatants (SN) 24 h after infection with adenoviruses encoding DsRed (control), wild-type CRELD2 (WT), CRELD2 lacking its ER retention signal (ΔREDL) or CRELD2 in which RDEL was replaced by KDEL (KDEL). b, Recovery 16 h after scratch injury of adenovirus-infected HCAEC monolayers. Exemplary fluorescence microscopy images after BCECF-AM staining; the positions of the initial scratches are indicated by dashed lines (scale bar, 250 μm). Summary data from eight to nine experiments. *P < 0.05; ***P < 0.001 (one-way ANOVA with Dunnett test). c, MI was induced in Tie2-GFP mice. After 3 days, tissue samples from the infarct region were explanted and infected with adenoviruses encoding DsRed (control), wild-type CRELD2, CRELD2-ΔREDL or CRELD2-KDEL. Exemplary fluorescence microscopy images (scale bar, 100 μm) and average sprout length after 3 days. There were 7–10 experiments. *P < 0.05; **P < 0.01 (one-way ANOVA with Dunnett test). d, Recovery after scratch injury of HCAEC monolayers stimulated for 16 h with various concentrations of recombinant CRELD2. Dashed lines indicate recovery in unstimulated cells and VEGFA (50 ng ml⁻¹)-stimulated cells. There were 5–11 experiments. Data are presented as mean values ± s.e.m. EC₅₀ denotes half maximal effective concentration as calculated by four-parameter logistic regression. e, EdU incorporation in HCAECs cultured for 24 h in the absence or presence of VEGFA (50 ng ml⁻¹) or CRELD2 (100 ng ml⁻¹). Exemplary fluorescence images (scale bar, 100 μm). Summary data are from six experiments. **P < 0.01 (one-way ANOVA with Dunnett test). f, HCAEC network formation after stimulation for 4 h with VEGFA (50 ng ml⁻¹) or CRELD2 (100 ng ml⁻¹). Exemplary images after BCECF-AM staining (scale bar, 500 μm). Summary data are from four to five experiments. **P < 0.01 (one-way ANOVA with Dunnett test). Unless otherwise stated, individual data points, mean values and s.e.m. are shown.

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