Verification of loss of BNip3 expression and re-expression

1. A, B BNip3 protein expression determined by immunohistochemistry at early carcinoma stage of tumorigenesis (65–80 days of age) in wild-type (A) and BNip3 null (B). Scale bar is 200 μm.
2. C Western blot analysis for expression of BNip3 in extracts from wild-type and BNip3 null MECs grown at 20% and 1% oxygen.
3. D Cell cycle phase distribution of wild-type and BNip3 null MECs measured in triplicate experiments by BrdU labeling over 5 h in culture followed by labeling of fixed cells with α-BrdU and PI. Results are expressed as the mean ± SEM. ****P < 0.0001.
4. E Western blot analysis for expression of exogenous BNip3 in extracts from parental BNip3 null MECs (Par), BNip3 null MECs expressing empty control vector (+ Empty vector) or BNip3 null MECs expressing exogenous BNip3 (+ BNip3-WT) grown at 20% and 1% oxygen.