Fig 6. Stat3 and Hif-1α cooperate in the acute response to oxidative stress.

A) Hif-1α expression in -/- WT MEFs transfected with shRNAs against Hif-1α and subjected to hypoxia for 2 h was monitored by qRT-PCR. Hif-1α was normalised against Hbs1l. Values are presented as mean ± SEM, n = 3 per group. *P<0,1, **P<0.01, ****P<0.0001. B) -/- WT MEFs transfected with shRNAs against Hif-1α were subjected to hypoxia for 2 h and Hif-1α protein levels were determined by SDS-PAGE and immunoblotting. C) -/-WT MEFs were transfected with shRNA vectors for Hif-1α or control vector and subjected to 2 h hypoxia (1% O₂) and 2 h reoxygenation (H/R). Isolated RNA was enriched for mRNA and analysed for Kcnb1 and Hbs1l expression by qRT-PCR. Kcnb1 expression was normalised to Hbs1l. Data are expressed as mean ± SEM, n = 3. D) As in (C) except that cells were stimulated with 100 μM H₂O₂ for 1 h. E) MEFs were untreated (-), treated with 100 μM H₂O₂ for 1 h or hypoxia (1% O₂) for 2 h, as indicated (+) and stabilisation of Hif-1α was determined by SDS-PAGE and immunoblotting. F) MEFs were untreated (-) or subjected to hypoxia for 2 h (+). Isolated RNA was enriched for mRNA and analysed for Hif-1α and Hbs1l expression by qRT-PCR. Hif-1α expression was normalised to Hbs1l. Data are expressed as mean ± SEM, n = 3. G) MEFs were untreated (-), treated with H₂O₂ for 1 h or subjected to hypoxia for 4 h, as indicated. Isolated RNA was enriched for mRNA and analysed for Kcnb1 and Hbs1l expression by qRT-PCR. Kcnb1 expression was normalised to Hbs1l. Data are expressed as mean ± SEM, n = 3.