Immunohistochemical and Flow Cytometry Analysis
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Variable analysis
- Paraffin sections were stained with hematoxylin and eosin (H&E) or used for immunohistochemical staining
- Primary antibodies and detection systems
- Secondary antibodies conjugated with Alexa Fluor 488 or Alexa Fluor 594
- BrdU was injected i.p. into pregnant females 2 hr prior to embryo harvest
- Fluorescent images obtained using a Zeiss Axioplan2 microscope
- Flow cytometry performed using cells isolated from yolk sacs and lungs
- RNA preparation and qRT-PCR analysis of purified cells
- Slides were counterstained with DAPI (Vector Lab)
- Antibodies used for flow cytometry are listed in Supplemental Materials
- Annexin V kit was from eBioscience
- Immunohistochemical staining as described in references 26 and 27
- No explicit negative controls mentioned
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