![Figure 4. Stimulation of human U937 monocytes with anti-MyD-1 stimulates PI 3-kinase. (A) Uninduced [32P]-labeled U937:Δp85 cells were stimulated in the presence (▪) and absence (□) of anti-MyD-1 (10 μg/mL) for the indicated times at 37°C. Pretreatment (30 minutes) with wortmannin (1 nM) had little effect on basal PIP3 levels (▵) but suppressed anti-MyD-1-stimulated PIP3 production (▴). Similar anti-MyD-1 stimulations were carried out on cells induced to overexpress the dominant-negative form of p85. IPTG-induced U937:Δp85 cells were stimulated in the presence (•) and absence (○) of anti-MyD-1 (10 μg/mL) for the indicated times at 37°C. Lipids were extracted and [32P]PIP3 production was identified by TLC and quantified by liquid scintillation counting. Data are presented as means ± SD, n = 3, and are representative of 5 independent experiments demonstrating coupling of MyD-1 to PI 3-kinase. (B) Anti-SHP-2 immune complexes from U937 cells were assessed for p85 expression, tyrosine phosphorylation, and SHP-2 expression.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/102/7/10.1182_blood-2002-11-3596/6/h81934992004.jpeg?Expires=1722602993&Signature=WLkCRT8tx-m~eKu9XUKf8~TPwMdUTxdrY6bSQOrw4PBh29nsljQI5R0ofp1~TDtnln4sJy6Cpa5y3kv3IxDptbriM7XfdmhyMvwGKP-2W-GmHpE9~ltC6xqNZi7vDr~9oswyDg2hrGO4Dk6NJpSTYv6~tGMeAR49eDNPQoUAee5M7iE4hQPbpvo6bJYeISIkdU4xaCF8-jxvspQkWd5aQSunaRQohhTNHSfDhg4Fs1mazk0NR4WWw981a2lYSy5T~RY7I-Az2rfgvJvTdHxTu2FsdUWJdgtlppyJ~6fVMFYDl8bBwnLLjetm8f0n8r7O~BYZWFU~MR4o9nGN2sQqYA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Stimulation of human U937 monocytes with anti-MyD-1 stimulates PI 3-kinase. (A) Uninduced [32P]-labeled U937:Δp85 cells were stimulated in the presence (▪) and absence (□) of anti-MyD-1 (10 μg/mL) for the indicated times at 37°C. Pretreatment (30 minutes) with wortmannin (1 nM) had little effect on basal PIP3 levels (▵) but suppressed anti-MyD-1-stimulated PIP3 production (▴). Similar anti-MyD-1 stimulations were carried out on cells induced to overexpress the dominant-negative form of p85. IPTG-induced U937:Δp85 cells were stimulated in the presence (•) and absence (○) of anti-MyD-1 (10 μg/mL) for the indicated times at 37°C. Lipids were extracted and [32P]PIP3 production was identified by TLC and quantified by liquid scintillation counting. Data are presented as means ± SD, n = 3, and are representative of 5 independent experiments demonstrating coupling of MyD-1 to PI 3-kinase. (B) Anti-SHP-2 immune complexes from U937 cells were assessed for p85 expression, tyrosine phosphorylation, and SHP-2 expression.
