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Shaking can cause foaming, which indicates that the peptide chains are breaking and the compound is losing its efficacy for your metabolic assays

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We also thank Drs Tobias Bruegmann, Eicke Latz and Philipp Sasse from University of Bonn for scientific discussions, Dr Fernando Pereira de Almeida from Federal University of Rio de Janeiro for assistance with microscopy image acquisition, Sandro Torrentes da Cunha from Federal University of Rio de Janeiro for assistance with histology preparations and Dr Bernardo Tura from Brazilian National Institute of Cardiology for assistance with the statistical analyses

Annals of Thoracic Surgery 114(1):327-333, 2022

After 48 h of growth in NGF or BDNF, cultures were either maintained with trophic support or deprived of it, in the absence or presence of each specific compound for the entire deprivation period