![Fig. 1. Activation of THP-1 cells with MoAb My43 and huIgA induces tyrosine phosphorylation of cellular proteins. THP-1 cells (5 × 105/sample) were activated at 37°C for 5 minutes by the addition of 5 μg/mL Mo2 (anti-CD14, lane 1), My43 cell culture supernatant 1:2 (anti-CD89, lane 2), 100 μg/mL huIgA (lane 3), 100 μg/mL huIgA + 100 μg/mL F′2RAhuIgA (lane 4), 100 μg/mL F′2RAhuIgA (lane 5), or NC particles (1 mg/sample) incubated in PBS containing different concentrations of huIgA, 3 mg/mL (lane 6), 1 mg/mL (lane 7), 0.33 mg/mL (lane 8), 0.11 mg/mL (lane 9), and 0.037 mg/mL (lane 10), and NC particles incubated in PBS containing 5% BSA only (lanes 11 and 12). Cellular proteins were isolated, separated by SDS-PAGE, and transferred to NC sheets as described in the experimental section. Immunoblotting of tyrosine-phosphorylated proteins was performed with the MoAb 4G10 followed by development with ECL as described (A). The immunoblot was then stripped and reprobed with a rabbit polyclonal anti-Syk antibody (lanes 1 through 11 in [B]) or control rabbit polyclonal IgG (lane 12 in [B]).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/91/2/10.1182_blood.v91.2.383/3/blod4024201.jpeg?Expires=1735545253&Signature=VryR67O8ncObiS66IEW~Uu56cacNTv5DxxZkC9qsfZ7V~r-IJmWCamfhqH5S~GI~ZdgHOuJHnqlSV5C8S2PWQb35or~2Ao~cvk2Reh~k7UD60RA7Z7i2-cQCeaeRtbpYLShm~7Fn7nZgGuT2gK7TdWjD~vl1iHqYdpaLUhJ1RQWGD1xrn4BPOUrcvWlH4Qlqb3lk3kz-hfE7-4W7QxDhqOAr~KFDKGHvlYd32HLZd3eWEg1UsEWhS-SFXnNyRPJ8s9~ksJxoVpnm2mjuppPpDNa3cGV3KVdkU~X7gn8ziGS40FphOlL~RzPrQ66bDK7xNCOkkX~lPlgouYdyQvA5-Q__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Activation of THP-1 cells with MoAb My43 and huIgA induces tyrosine phosphorylation of cellular proteins. THP-1 cells (5 × 105/sample) were activated at 37°C for 5 minutes by the addition of 5 μg/mL Mo2 (anti-CD14, lane 1), My43 cell culture supernatant 1:2 (anti-CD89, lane 2), 100 μg/mL huIgA (lane 3), 100 μg/mL huIgA + 100 μg/mL F′2RAhuIgA (lane 4), 100 μg/mL F′2RAhuIgA (lane 5), or NC particles (1 mg/sample) incubated in PBS containing different concentrations of huIgA, 3 mg/mL (lane 6), 1 mg/mL (lane 7), 0.33 mg/mL (lane 8), 0.11 mg/mL (lane 9), and 0.037 mg/mL (lane 10), and NC particles incubated in PBS containing 5% BSA only (lanes 11 and 12). Cellular proteins were isolated, separated by SDS-PAGE, and transferred to NC sheets as described in the experimental section. Immunoblotting of tyrosine-phosphorylated proteins was performed with the MoAb 4G10 followed by development with ECL as described (A). The immunoblot was then stripped and reprobed with a rabbit polyclonal anti-Syk antibody (lanes 1 through 11 in [B]) or control rabbit polyclonal IgG (lane 12 in [B]).
