Fig. 6

ADORA2B knockdown inhibits HIF-1α, Akt, and ERK1/2 activation. Immunoblotting analysis of ADORA2B, HIF-1α, pERK1/2, and pAkt proteins in the Mock and shADORA2B cells under both normoxic and hypoxic conditions. The expression of ADORA2B, HIF-1α, pERK1/2, and pAkt protein were up-regulate in the shMock cells of hypoxia compared with a normoxia. The HIF-1α protein levels in the shADORA2B-transfected cells (HSC-3- and -4-derived transfectants) also have decreased markedly compared with that in the shMock-transfected cells under both normoxic and hypoxic conditions. ADORA2B knockdown causes decreased levels of pAkt and pERK1/2 compared with the shMock-transfected cells (HSC-3- and -4-derived transfectants); the AKT and ERK1/2 protein levels are unchanged under both normoxic and hypoxic conditions. Densitometric ADORA2B, HIF-1α, pAkt/Akt and pERK/ERK protein data were normalized to the α-tubulin protein levels