Phosphorylation of p47phox in EBV-transformed B lymphocytes from p67phox-deficient patients and from healthy donors. (A) EBV-transformed B lymphocytes from healthy donors (EBV-L) and p67phox-deficient patients (EBV-l-p67phox^−/−) (15 × 10⁶/400 µL) were stimulated with PMA (200 ng/mL) for 10 minutes and the reaction was stopped by denaturation with 100 µL Laemmli sample buffer (5×). Homogenates were subjected to SDS-PAGE and western blotting using anti-gp91phox, anti-p67phox, and anti-p47phox antibodies. (B) EBV-transformed B lymphocytes from healthy donors (EBV-L) and p67phox-deficient patients (EBV-l-p67phox^−/−) (15 × 10⁶/400 µL) were stimulated with PMA (200 ng/mL) for 10 minutes and the reaction was stopped by denaturation with 100 µL Laemmli sample buffer (5×). Homogenates were subjected to SDS-PAGE and western blotting using different anti-phospho-p47phox antibodies: anti-phospho-Ser304, anti-phospho-Ser315, anti-phospho-Ser320, anti-phospho-Ser328, and anti-phospho-Ser-345 antibodies and anti-p47phox antibody. Experiments were repeated 3 times.