10.1002/cmdc.202000763
ChemMedChem
FULL PAPER
48 h using a 10X objective. Ten random micrographs per well were
obtained and migration area was quantified using NISElements 3.0 (Nikon)
software. Wound closure measurements were normalized to the maximum
scratch area.
Conflict of interest
The authors declare no conflict of interest.
Keywords: Rac1 inhibitors • N,N´-disubstituted guanidines •
protein-protein interactions inhibitors • lung cancer
Indirect immunofluorescence assay (IFI): The procedure for IFI was
performed as previously described.[52] Subconfluent A549 cells grown on
glass coverslips in 24-well plates were fixed with methanol for 10min at
−20 °C. After three washes with PBS, the coverslips were inverted on a
drop of diluted primary antibody for 30 min at 37 °C, and then returned to
culture dishes and subjected to three additional washes with PBS.
Afterwards, cells were incubated with diluted secondary antibody for 30
min at 37 °C. Nuclei were counterstained with DAPI (4 μg/ml in PBS) for 5
min at room temperature protected from the light. Finally, coverslips were
rinsed, mounted and photographed with an Olympus BX51 with
epifluorescence optics.
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This work was partly supported by Agencia Nacional de
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Investigación Científica y Tecnológica (FONCyT) PICT-201-0362
to MJC and PICT-2018-1036 to JB, Consejo Nacional de
Investigaciones Científicas y Técnicas (CONICET) PIP2014-2016
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The manuscript was written through contributions of all authors.
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