Fig. 3
From: Cellular direct conversion by cell penetrable OCT4-30Kc19 protein and BMP4 growth factor
Morphological, phenotypical, and genomic changes in HUVECs after OCT4-30Kc19 and BMP4 treatment. (A) Morphological changes in HUVECs after OCT4-30Kc19 and BMP4 treatment. Stimulation of HUVECs with 40 μg/ml of OCT4-30Kc19 and 10 ng/ml of BMP4 for 48 h resulted in elongated morphology in HUVECs. Cell morphology was visualized with rhodamine-phalloidin staining. Blue, red represent the nucleus and actin structure, respectively. Scale bar, 20 μm. (B) Phenotypical changes in HUVECs after OCT4-30Kc19 and BMP4 treatment. Immunofluorescence images of protein-treated HUVECs showed elevated expression of CD31 and αSMA. Anti-CD31-antibody (1:200) and anti-αSMA-antibody (1:200) were used as primary antibodies. Blue, green, red represent the nucleus, CD31, and αSMA, respectively. Scale bar, 20 μm. (C) Genomic changes in HUVECs after OCT4-30Kc19 and BMP4 treatment. Quantitative polymerase chain reaction (qPCR) analysis for endothelial, mesenchymal, and endothelial-mesenchymal transition (EndMT) markers showed an elevation in fold induction after both protein treatments (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001. (D) Tube formation assay to evaluate the effects of OCT4-30Kc19 and BMP4 on angiogenesis. Tube formation of HUVECs was visualized with phalloidin staining after OCT4-30Kc19 and BMP4 treatment. Scale bar, 500 μm. (E) Quantitative analysis of branches per field (n = 3). ***p < 0.001