IL-7 Induces an Epitope Masking of γc Protein in IL-7 Receptor Signaling Complex

Figure 4

The γc epitope masking by the conformational change of γc. T cells were stimulated in the presence or absence of IL-7 for 10 minutes. (a) γc protein level was analyzed by immunoblot. (b) γc and IL-7Ra mRNA level were measured by real-time PCR. Blots are representative of four independent experiments. Data are summary of four independent experiments (mean and SEM). (c) LNT cells were isolated from WT and γc^−/−γcTg mice and stimulated with IL-7 or medium. The γc (4G3) expression was analyzed 10 minutes after stimulation. Data are summary of three independent experiments (mean and SEM). (d) T cells were immunostained with 4G3, TUGm2, 3E12, and anti-γc pAb 10 minutes after IL-7 stimulation. The maximum value of -axis is 250 for all histograms. Histograms are representative of four independent experiments. (e) The relative γc expression was calculated as the ΔMFI associated with T cells incubated at the indicated condition over the ΔMFI associated with fresh T cells. Data are summary of four independent experiments (mean and SEM). (f) Confocal microscopic analysis of surface γc protein. Murine LNT cells were stimulated with IL-7 for 10 and 30 minutes. The fresh or stimulated cells were fixed and stained with biotin-conjugated TUGm2 and pAb. And Streptavidin Alexa Fluor 594 was used to detect surface γc protein (red) and DAPI was used to stain nuclei (blue). Data are representative of three experiments.

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