Cytoplasmic localisation of p53 in proliferating ES cells. R1, D3 and CGR8 mouse embryonal stem cells were plated on cover slips (R1 and D3 in the presence of feeders, CGR8 in the absence of feeders). Two days after plating, cells were fixed in acetone/methanol, permeabilized with Triton-X-100 and incubated with the anti-p53 antibody Pab 421. After incubation, cover slips were washed and incubated with an antibody directed against mouse IgG, coupled to Alexa-Fluor-488 (green). To visualize the nuclei, cover slips were incubated with Draq5 (blue). (B) R1 ES cells were transfected with RGC-lacZ or with a vector control together with GFP to visualize transfection efficiency. Two days after transfection, cells were fixed with paraformaldehyde and incubated with X-gal in reaction buffer to monitor β-galactosidase expression. Arrows point to lacZ expressing R1 cells.
Solozobova et al. BMC Cell Biology 2009 10:46 doi:10.1186/1471-2121-10-46