Anti-CD20/CD74 HexAbs inducing activation of ERK and JNK MAP kinases and inhibiting the Akt and NFκB signaling. (A) Rapid and sustained phosphorylation of ERK1/2 and JNK MAP kinases. JeKo-1 cells were treated with 10-nM concentrations of 74-(20)-(20) (top panel) or 20-(74)-(74) (middle panel) or with a combination of parental mAbs (bottom panel) for the indicated time points up to 24 hours. Cells were lysed, and samples were evaluated after probing with relevant Abs. β-actin was used as the loading control. (B) The CD20-targeting HexAbs, like veltuzumab, inhibited the phosphorylation of c-Src⁴¹⁶. (C) HexAbs inhibited the nuclear translocation of p65. JeKo-1 cells were treated with the indicated Abs for 72 hours, and nuclear and cytosolic extracts were prepared. Equal amounts of proteins were evaluated for the expression levels of p65 in cytosolic and nuclear fractions. Brg-1 (nuclear) and β-actin (cytosolic) served as loading controls. (D) HexAbs inhibited the phosphorylation of Akt. JeKo-1 cells were treated with the indicated Abs for 24 hours, lysed, and probed for pAkt (serine 473).