Journal: The Journal of Experimental Medicine

Article Title: Impaired Interleukin 4 Signaling in T Helper Type 1 Cells

doi:

Figure Lengend Snippet: IL-4R expression in Th1s and Th2s. ( A ) 10 5 two-round– primed TCR transgenic Th1s and Th2s were incubated with the M1 anti–IL-4R monoclonal antibody or a rat isotype control in the presence or absence of IL-4 (1.5 ng/ml). The cells were washed and stained with FITC-labeled Fab goat anti–rat IgG. Samples were analyzed with a FACSCAN ® . ( B ) To calculate the ratio of IL-4R expression in Th1s and Th2s, the mean fluorescence intensity (MFI) of M1-stained Th1s minus the MFI of isotype control Th1s was divided by the MFI of M1-stained Th2s minus the MFI of isotype control Th2s.

Article Snippet: FITC-labeled goat Fab′ fragment against rat IgG (Southern Biotechnology Associates, Birmingham, AL) were added to stained cells for 20 min. For CD30 staining, cells were first blocked with 10 μl of 2.4G2 rat anti–mouse FcγRII/III ascitic fluid for 5 min before staining with PE-labeled monoclonal antibody against mouse CD30 ( PharMingen , San Diego, CA) for 20 min in FACS ® buffer.

Techniques: Expressing, Transgenic Assay, Incubation, Staining, Labeling, Fluorescence