Figure 6.

CF Null Eosinophil Granularity Can Be Stabilized by Inhibition of Cysteine Protease Activity

(A–C) Protease inhibitors were added to in vitro cultured eosinophils after 4 day growth in IL-5 and replenished every other day for 9 days. Histogram overlays show increase in SSC of Cst7^−/− eosinophils treated with (A) recombinant mouse CF or with the irreversible broad-spectrum cysteine protease inhibitors z-FA-fmk (B) and E64d (C).

(D) Quantitation of SSC-rescue with CF and z-FA-fmk. Data are expressed as percent restoration of the difference in mean SSC between control WT & Cst7^−/− eosinophils; means ± SD of three independent experiments are shown.

(E) Lysates from 200,000 cells cultured as in (D) were separated by reducing SDS-PAGE (z-FA-fmk = 0.4 μM). MBP, Epx, CF, and GAPDH loading control were detected by immunoblot. Data are representative of at least three independent experiments.