Fig. 1.

Endogenously synthesized β2m is not required for open conformer-dependent signaling in DAOY cells. a DAOY cells were treated with either β₂m-specific siRNA or random siRNA control. Exogenous β₂m was added 48 h later and lysates were subjected to the Western blot analysis using β2m- or GAPDH-specific antibodies, as indicated. b DAOY cells treated in the same manner as in a were stained with W6/32 (green lines; right), HC-10 (blue lines; middle) monoclonal antibodies, or secondary antibody alone (red lines; left), and analyzed by flow cytometry. c Exogenous β₂m in the absence or presence of W6/32 (W) or HC-10 (H) monoclonal antibodies was added to DAOY cells pretreated with β₂m-specific siRNA, as indicated. Cell lysates were probed by β₂m-, GAPDH-, phospho-ERK1/2, or total ERK1/2-specific antibodies. C-control cells treated with neither siRNA nor exogenous β2m