Fig 1: Angiogenic protein profiles of CM, EV-depleted CM, and lysed and non-lysed EVs of DPSCs and BM-MSCs. A screening for 55 pro- and anti-angiogenic factors was performed on EV-depleted CM and lysed EVs derived from DPSCs and BM-MSCs (n = 3, 3 different donors). Relative pixel density was expressed as percentage after subtraction of the negative control and normalization to positive reference spots (indicated as ‘-’ and ‘+’) and displayed in a heat map (A). Representative antibody blots are presented for each condition (B), with the corresponding protein locations (C). Semi-quantitative results of proteins angiopoietin-1 (Angpt-1) (D), IGFBP-1 (E), IGFBP-3 (F), VEGF (G) (n = 5, 5 different donors (DPSCs) or 5 different isolations of 3 different donors (BM-MSCs)) and TIMP-1 (H) (n = 7, 7 different donors (DPSCs) or 7 different isolations of 3 different donors (BM-MSCs)) were confirmed with ELISA on CM, EV-depleted CM and (lysed) EVs of both stem cell types (factors indicated in red). Limited or no expression of these angiogenic factors could be detected in non-lysed EV samples, suggesting that the EV-associated factors are located intravesicularly (I, nd = not detectable). Significant differences in angiogenic protein content between stem cell types and secretion fractions could be observed as determined with mixed model ANOVA. Quantitative data are shown as mean ± S.E.M. * p < 0.05, ** p < 0.01, *** p < 0.0001 between EVs, CM and EV-depleted CM, # p < 0.05, ## p < 0.01, ### p < 0.0001 between DPSCs and BM-MSCs.
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