1. Supplementary Figure 1 A B C SW620 HT29 SW620
RT-PCR
Western Blot
1% O2 (h)
1% O2 (hr)
HT29
Nur77
18s rRNA
β -actin
SW620
Nur77
18s rRNA
β -actin
HT29 *
SW620 * *
HT29 *
SW620 * B
CoCl2 (h)
Nur77
β -actin
SW620
CoCl2 (h)
HCT116
β -actin
Nur77 C
DFX (h) *
SW480
Nur77
β -actin
DFX (h) *
HT29
Nur77
β -actin
Supplementary Figure 1. Hypoxic conditions induce the mRNA and protein lev­els of Nur77. Colon cancer cells were serum starved for 24 h, and the­n (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 mM) 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.