Fig. 2

Characterization of the growth of Wt and Vdac1 ^−/− MEF. a Both cell lines were seeded at the same density and incubated in Nx or Hx for the indicated number of days. The mean ± SEM is representative of four independent experiments carried out in duplicate. A p < 0.0002 shows significant difference from normoxia for the Vdac1 ^−/− MEF. b Immunoblotting of HeLa cells transfected with pFlag-VDAC1. c Immunoblotting and proliferation assay of MEF tranfected with pFlag-VDAC1 d Relative migration of Wt and Vdac1 ^−/− MEF in Nx as evaluated with a xCELLigence system. The mean ± SEM is representative of two independent experiments carried out in quadruplicate. e Characterization of the growth of Wt and Vdac1 ^−/− MEF incubated in Nx or in Hx for 3 days in the absence or presence of an inhibitor of MEK (U0126, 10 μM). The mean ± SEM is representative of two independent experiments carried out in duplicate. f Wt (+) and Vdac1 ^−/− (−) cells were incubated in Nx or Hx in the absence or presence of an inhibitor of MEK (U0126). Cell lysates were analyzed by immunoblotting for phospho-ERK (P-ERK), total ERK (ERK) and β-tubulin as a loading control. g Immunofluorescence of P-ERK in Wt and Vdac1 ^−/− MEF in Hx. h Wt (+) and Vdac1 ^−/− (−) MEF were incubated in Nx or Hx for 72 h and cell lysates were analyzed by immunoblotting for DUSP6. ARD1 was used as a loading control. (i) Immunoblotting of Wt MEF and Hif-1α^−/− MEF for DUSP6