B

C

A

Supplementary Figure 1

SW620

CoCl2 (h) 0 2 4 8 24

Nur77

β -actin

β -actin

Nur77HCT116

RT-PCR Western Blot

DFX (h) 0 2 4 8 24

SW480 Nur77

β -actin

HT29 Nur77

β -actin

HT29Nur77

18s rRNA

Nur77

β -actin

SW620Nur77

18s rRNA

Nur77

β -actin

HT29

* * **

SW620

*

* * *

**

**

** *

*

*

* * *

* **

*

**

* *

HT29*

** *

SW620

0 2 4 8 241% O2 (h) 0 2 4 8 241% O2 (hr)

CoCl2 (h) 0 2 4 8 24

DFX (h) 0 2 4 8 24

Supplementary Figure 1. Hypoxic conditions induce the mRNA and protein lev els of Nur77. Colon cancer cells were serum starved for 24 h, and the n (A) incubated at 1% O2 level in a modulator; or treated with the hypoxia mimetic (B) cobalt chloride (CoCl2, 100 mM) or (C) desferrioxamine (DFX, 100 M) for the time indicated. Total RNA or proteins were extracted from cell lysates for subsequent RT-RCR and Western blotting to determine the mRNA and protein expression respectively. Whole-cell lysates were analyzed by immunoblotting. β-actin was used as internal control for these analyses. Immunoblots were quantified by densitometry and the values were normalized with β-actin. All blots shown are representative of three independent experiments. bar, ± SD. *, P < 0.05.