It was performed as described in previous studies (22 (link)). Cells were incubated overnight at 4°C with primary antibody for p-H3 (ab14955 Abcam, 1/2000) and active caspase-3 (AF835 R&D Systems, 1/500). Then, cells were incubated for 1 hour with secondary Alexa Fluor 555 IgG anti-mouse (A21422 Invitrogen, 1/500) or anti-rabbit (A31572 Invitrogen, 1/500) antibodies at room temperature in darkness. For nuclear DNA staining, Hoechst 33342 (Sigma) was used. Pictures were taken with an Eclipse 80i microscope and processed with the NIS Elements Advances Research software (Nikon).
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Immunofluorescent Imaging of Apoptosis Markers
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Corresponding Organization : Ikerbasque
Other organizations : CIC bioGUNE, Instituto de Salud Carlos III, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, Albert Einstein College of Medicine, Osakidetza
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Variable analysis
independent variables
- Incubation of cells overnight at 4°C with primary antibody for p-H3 (ab14955 Abcam, 1/2000) and active caspase-3 (AF835 R&D Systems, 1/500)
- Immunostaining of p-H3 and active caspase-3
- Incubation of cells for 1 hour with secondary Alexa Fluor 555 IgG anti-mouse (A21422 Invitrogen, 1/500) or anti-rabbit (A31572 Invitrogen, 1/500) antibodies at room temperature in darkness
- Nuclear DNA staining with Hoechst 33342 (Sigma)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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