Skip to main content
. 2012 Jul 2;32(19):2463–2474. doi: 10.1038/onc.2012.256

Figure 3.

Figure 3

Expression and localization of ERBB2 receptor isoforms in HMLE cells. (a) Lysates from parental or recombinant ERBB2-transduced HMLE cells were probed with anti-ERBB2 antibody in a western blot assay. Actin was blotted as loading control. (b) Recombinant HMLE cells were fractionated into cytoplasmic ‘C', membrane ‘M' and nuclear ‘N' fractions, followed by western blotting for ERBB2. Control antibodies identify proteins restricted to the cytoplasm (GAPDH, actin), the plasma membrane (NaK-ATPase) and the nucleus (Lamin A/C). p110 HMLE also express the p95cyto isoform due to translation from the AUG codon corresponding to methionine 687. (c) Confocal microscopy of recombinant HMLE cells. Plasma membranes were stained with concanavalin A (green), followed by a mouse anti-ERBB2 antibody and anti-mouse Cy5 (red) for receptor labeling. p185, p110 and p95m reside primarily in the plasma membrane, whereas p95cyto and p95n reside in the cytoplasm and nucleus, respectively. Images are representative of the population at large and were taken at × 80 magnification on a Zeiss confocal microscope. Cells expressing p95n were analyzed at higher magnification to confirm nuclear localization.