Protein Expression Analysis of 3D-COL hPDLSCs

hPDLSCs were seeded at 80 × 10³/well in 6-well plates in basal medium alone or in the presence of 3D-COL, 3D-COL/CM, 3D-COL/EVs or 3D-COL/PEI-EVs. After 28 days in culture, cells were harvested and proteins were collected from 3D-COL/hPDLSCs, 3D-COL/hPDLSCs/CM, 3D-COL/hPDLSCs/EVs and 3D-COL/hPDLSCs/PEI-EVs samples (40 μg/sample). The western blot procedure was performed as previously reported (Ballerini et al., 2017 (link)). VEGFA (Santa Cruz Biotechnology; 1:1000) and VEGFR2 (Santa Cruz Biotechnology; 1:1000) were used as primary antibody. β-Actin (Santa Cruz Biotechnology; 1:750) was used to assess the uniform protein loading (Libro et al., 2016 (link)). Bands were analyzed by the ECL method using Alliance 2.7 (UVItec Limited, Cambridge, United Kingdom). Protein bands were quantified with a computer program (ImageJ software). Entire blots were visible in Supplementary Figure S1.

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Pizzicannella J., Gugliandolo A., Orsini T., Fontana A., Ventrella A., Mazzon E., Bramanti P., Diomede F, & Trubiani O. (2019). Engineered Extracellular Vesicles From Human Periodontal-Ligament Stem Cells Increase VEGF/VEGFR2 Expression During Bone Regeneration. Frontiers in Physiology, 10, 512.

Publication 2019

VEGFA VEGFR2 β-Actin Alliance 2.7

Actin Antibody Cells Protein Vegfr2 Western blot

Corresponding Organization :

Other organizations : University of Chieti-Pescara, Centro Neurolesi Bonino Pulejo, Institute of Cell Biology and Neurobiology, National Research Council

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Variable analysis

independent variables

3D-COL
3D-COL/CM
3D-COL/EVs
3D-COL/PEI-EVs

dependent variables

VEGFA protein expression
VEGFR2 protein expression

control variables

HPDLSCs seeded at 80 × 10^3/well in 6-well plates
Basal medium
Protein loading (β-Actin)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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