Figure 1
From: Regulation of membrane ruffling by polarized STIM1 and ORAI1 in cortactin-rich domains
Immunolocalization of phospho-STIM1. C2C12 cells were plated onto collagen-coated glass coverslips, and fixed after 48 h in culture. Immunolocalization of phospho-STIM1 was performed for the three ERK1/2 target sites (pS575, pS605, and pS621) using phospho-specific antibodies and anti-sheep IgG labelled with Alexa Fluor 594 as a secondary antibody. Negative controls were performed by incubating cells with 0.5 μM PD0325901 for 20 min before fixation. Additional negative controls were performed, treating fixed cells with lambda phosphatase (10 U/μl) overnight at 30 °C. Total STIM1 was immunolocalized using an anti-STIM1 antibody and anti-rabbit IgG as a secondary antibody. Images are representative of at least 6 independent experiments. Bar = 10 μm.
