Internalization of B Cell Receptors in Human EU12 μHC+ Immature B Cells Specifically Alters Downstream Signaling Events
Figure 4
Internalization of BCR results in delayed activation of SYK and sustained activation of ERK1/2 in EU12 μHC+ cells upon restimulation. EU12 μHC+ cells were directly stimulated with goat anti-IgM antibody F(ab′)2 fragments (10 μg/mL) or pretreated with goat anti-IgM antibody F(ab′)2 fragments (2 μg/mL) for 30 min, washed, recovered, and restimulated upon goat anti-IgM antibody F(ab′)2 fragments (10 μg/mL). Cells were collected at 0, 1, 3, 10, 15, and 30 min and analyzed for CD19, SYK, and ERK phosphorylation. The membranes were stripped and blotted with anti-CD19, anti-SYK, anti-ERK, or anti-β-ACTIN antibodies as controls.
