Figure 4

Degranulation reaction is normal in Arf1-deficient BMMCs. (a) Either control (ctrl; n = 3) or Arf1-KO (n = 3) BMMCs at 5-week after culture sensitized with anti-DNP IgE were stimulated without (dotted line) or with (solid line) 20 ng/mL of DNP-BSA for 20 min and evaluated for pS6 (left) and pErk (right) signals by FACS. Shown are representative FACS profiles (top) and mean fluorescence intensities (MFI) (bottom). Gray thin lines on FACS plot indicate negative stained signal with isotype controls. Mean ± SD. (b) Control (ctrl) or Arf1-KO BMMCs sensitized with anti-DNP IgE were stimulated without (untreated) or with (+ DNA-BSA) 20 ng/mL of DNP-BSA for 20 min. Shown are FACS profiles and proportion of CD107a⁺ cells representative of four. (c) Proportions of CD107a⁺ cells in control (ctrl) or Arf1-KO BMMCs stimulated as in (b) for the indicated time periods were evaluated by FACS (n = 4, each). (d) Control (ctrl) or Arf1-KO BMMCs sensitized with anti-DNP IgE were stimulated with the indicated concentrations of DNP-BSA for 24 h (d; n = 5, each). Concentrations of IL-6 in the culture supernatants were then quantified by ELISA. (e) Control BMMCs sensitized with anti-DNP IgE were stimulated with 20 ng/mL of DNP-BSA along without (none) or with 0.1 µM of rapamycin (+ Rap), 0.1 µg/mL of FIPI (+ FIPI), or 0.1 µM of BFA (+ BFA) for 24 h (e; n = 9, each). Concentrations of IL-6 in the culture supernatants were then quantified by ELISA. **p < 0.01.