Figure 2.

RNA blot analyses of DMT1 expression in duodenum of iron-deficient mice. Three micrograms of poly(A)⁺ RNA from duodenum of a nontransgenic FVB/N control (C, lanes 1, 3, 5) and a transgenic H⁺/K⁺-ATPase β subunit (−1035 to +24)/DT-A iron-deficient littermate (DT-A, lanes 2, 4, 6) were electrophoresed in triplicate and blotted. Lanes 1 and 2 were hybridized with a ³²P-labeled coding-sequence probe [DMT1(total)] to detect all DMT1 transcripts. Lanes 3 and 4 were hybridized with an oligonucleotide probe [DMT1(IRE)] specific to DMT1 transcripts with an IRE, and lanes 5 and 6 were hybridized with an oligonucleotide probe [DMT1(non-IRE)] specific to the DMT1 splice-variant transcripts without an IRE. Blots were exposed to film for 8 h with an intensifying screen and rehybridized with a probe for GAPDH. Positions of RNA size markers are on the left.