Figure 3: Hp:Hb scavenging-dependent Akt phosphorylation. (a) Representative Western blot showing phosphorylated Akt (pSer473) and total Akt in lysates from macrophages stimulated with Hp1-1:Hb (1 mg/mL; HH in legend) for 20 minutes in the presence and absence of blocking anti-CD163 antibody clone RM3/1 (20  g/mL; RM3/1 in legend), colchicine (2.5  mol/L; colch in legend), or phosphoinositol 3-kinase inhibitor Ly294002 (12.5  mol/L; Ly in legend). (b) Quantitative ELISA determination of Akt (pSer473) phosphorylation in cell lysates from macrophages treated as in (a) above. (c) Comparison of Hp2-2:Hb versus Hp1-1:Hb scavenging on Akt phosphorylation in cell lysates, as measured by pSer473Akt-specific ELISA. Results in (b) and (c) are expressed as pSer473Akt (U/mg) ± SEM from three experiments. , .