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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: J Neurooncol. 2013 Oct 27;116(1):89–97. doi: 10.1007/s11060-013-1283-3

Fig. 1. PTEN-PI3K signaling is required for HIF1α accumulation in hypoxic conditions.

Fig. 1

(A) Immunoblot confirming expression of PTEN in reterovirally reconstituted U373MG-PTEN and U87MG-PTEN cell lines while parental U373 MG and U87MG cell lines lack its expression. (B) Muristerone induced U87 PTEN, G129E and G129R cell lines induced with 0.5μmol/L of muristerone were either serum starved or kept in 2% serum or 10% serum conditions for 36 hrs and were given normoxia (21% O2) or hypoxia (1% O2) for 4 hrs followed by preparation of nuclear extracts (for HIF1α) as well as whole cell extracts. (C) GFAPV12Ras PTENfl/fl and GFAP V12 Ras PTENfl/fl cre+ cell lines were serum starved or kept in 2% serum or 10% serum conditions for 4 hrs and were kept in normoxic (21% O2) or hypoxic (1% O2) conditions for 4 hrs followed by preparation of nuclear extracts (for HIF1α) as well as whole cell extracts. (D) U87MG glioma cell line treated with 10, 25 and 50μM SF1126 inhibited HIF1α accumulation under hypoxic conditions. Cells were treated with different concentrations of inhibitor for 30 min and then placed under normoxic or hypoxic conditions (1% O2) for 4 hours followed by the preparation of cell lysates for Western blot analysis of HIF1α.