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Cell nucleofection

JM Juan A. Montero
CS Cristina Sanchez-Fernandez
CL Carlos I. Lorda-Diez
JG Juan A. Garcia-Porrero
JH Juan M. Hurle
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The morphological evolution of interdigital cells positive for γH2AX was analyzed in cultures of dissociated interdigit mesoderm. For this purpose, cells were transfected with a construct encoding the human Btg2 gene cloned into the pCMV6-XL5 vector (SC115914, Origene). The efficiency of electroporation was confirmed using qPCR and immunohistochemistry. After two days of culture, cells were fixed in PFA and labeled with anti-γH2AX in combination with rhodamin-phallodin. The samples were examined using a confocal microscope to correlate changes in the distribution of the actin cytoskeleton with the intensity of γH2AX immunolabeling.

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