Immunofluorescence Staining of Testicular Cells

Immunofluorescence on sections and on surface-spread testicular cells were performed as previously described.^{15 (link), 65 (link)} Antibody against DOT1L (ab-64077 from Abcam), H3K79me2 (ab-3594 from Abcam) or AcH4 (06-866 from Millipore) were diluted 1/100 to 1/200. Pictures were taken with an Olympus BX63 microscope. Quantification was performed on pictures obtained from six samples per genotype using ImageJ 1.48v (http://imagej.nih.gov/ij/). T-tests were performed with GraphPad Prism 5.02.

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Moretti C., Serrentino M.E., Ialy-Radio C., Delessard M., Soboleva T.A., Tores F., Leduc M., Nitschké P., Drevet J.R., Tremethick D.J., Vaiman D., Kocer A, & Cocquet J. (2017). SLY regulates genes involved in chromatin remodeling and interacts with TBL1XR1 during sperm differentiation. Cell Death and Differentiation, 24(6), 1029-1044.

Publication 2017

ab-64077 ab-3594 BX63 microscope GraphPad Prism 5.02

Antibody Cells Dot1l Genotype Immunofluorescence Microscope Prism

Corresponding Organization :

Other organizations : Institut Cochin, Sorbonne Paris Cité, Université Paris Cité, Délégation Paris 5, Sorbonne Université, Australian National University, Maternité Port Royal, Centre Hospitalier Sainte-Anne, Institut des Maladies Génétiques Imagine, Assistance Publique – Hôpitaux de Paris, Genetique Reproduction and Developpement

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Variable analysis

independent variables

Genotype

dependent variables

DOT1L expression
H3K79me2 levels
AcH4 levels

control variables

Tissue samples (sections and surface-spread testicular cells)
Antibody concentrations (1/100 to 1/200 dilution)
Microscopy (Olympus BX63)
Image analysis software (ImageJ 1.48v)
Statistical analysis software (GraphPad Prism 5.02)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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