Figure 7
Immunofluorescent staining of ErbB1-4 receptors, phospho-ErbB2 and phospho-ErbB4 receptors. +SA mammary tumor cells were initially plated at a density of 2 × 105 cells/well in 6-well plates (3 replicates/group) and allowed to attach to the plates followed by incubation in mitogen-free media overnight. Afterwards, cells were stimulated with 100 ng/ml EGF for 10 min. The cells were fixed with methanol and incubated with primary antibody directed against specific ErbB receptors, and then incubated with rhodamine conjugated secondary antibody, as described in the Methods section. Magnification is 200×.