Localization of RARα and RARγ in Mouse Embryonal Carcinoma and Mouse Embryonic Stem Cells
Vitamin A is a precursor for retinoic acid (RA), a growth factor that positively influences embryonic development and cell differentiation. Addition of RA to mouse embryonic stem cells (mESC) and mouse embryonal carcinoma cells (mEC) induces neurogenesis presumably by complexing with nuclear RA receptors (RAR) that mediate gene expression. The goal of this study was to determine the specificity of 3 RARα and 1 RARγ antibodies and to localize RARα and RARγ protein expression in mESC and mEC cells. P19 mEC and R1ES cells were grown in undefined media supplemented with 15% fetal bovine serum (FBS). C57/BL6 ES cells were grown in chemically defined media. Fixed cells were reacted with RARα and RARγ antibodies using immunofluorescent protocols. Negative controls showed no reactivity. P19 mEC and both mESC cell types showed positive nuclear staining with both antibodies. Staining was variable in intensity throughout mEC and mESC colonies, suggesting variability in expression of the proteins. Incubation with RA for 24 to 72 hours did not increase or decrease nuclear expression. The results confirm the specificity of RARα 115 and RARγ 453 antibodies. The results also provide the first report showing localization of RARα and RARγ proteins in P19 mEC and two strains of mESC before and after incubation with RA.
FaMitah QuiAnna Buchanan,
"Localization of RARα and RARγ in Mouse Embryonal Carcinoma and Mouse Embryonic Stem Cells"
ETD Collection for Tennessee State University.