(A) NIH-3T3 cells expressing EGFRWT were infected with
lentiviral constructs to stably overexpress GGA2 (GGA2 OE) or GFP leading to
GGA2 overexpression in the former as indicated by Western blot. (B) GGA2 OE and
GFP control cells were seeded in serum starved media before being subjected to
20uM cycloheximide for 30 minutes. Lysates were harvested for time point 0.
Remaining cells were treated with 30ng/mL EGF and harvested at 1, 3, 6, 9, 12,
24 and 48 hours (h). Lysates were subjected to immunoblot analyses demonstrating
that EGFR and pEGFR levels were more persistent in cells overexpressing GGA2.
(C) The same experiment as described in (B), with 3ng/mL EGF also indicating
increased EGFR stability and activity in the setting of GGA2 overexpresion.