The p85 Regulatory Subunit of PI3K Mediates cAMP-PKA and Insulin Biological Effects on MCF-7 Cell Growth and Motility
Figure 2
Role of phosphorylation of S83 in the subcellular localization of p85α and IRS-1. MCF-7 cells transiently transfected with p85WT or p85A or p85D were seeded on 12 mm glass coverslips, with serum starved for 6 h and treated with 10 nM insulin for 10 and 20 minutes. As internal control cells were treated for 30 minutes with FBS. Immunofluorescence staining was performed as described in Materials and Methods. After incubation cells were fixed, permeabilized, and stained with anti-FLAG and anti-IRS-1 antibodies. Pearson’s coefficient is shown in each panel, overlapping images and Van Steensel’s cross-correlation functions (CCFs) were calculated for each experimental point (see Figure S1B). Transfection efficiency was determined by flow cytometry using anti-FLAG antibodies (see Section 2) and is shown in Figure S3B.
